Antibody to Parvalbumin - calcium binding protein used as a marker for an inhibitory interneuron subtype. Part of the ValidAb™ range of highly validated, data-rich antibodies.
Figure 1. Parvalbumin and Calretinin expressing interneurons in hippocampal CA1.
A section of hippocampal CA1 stained by HB6457 for Parvalbumin (mouse monoclonal) and HB6494 for Calretinin (rabbit polyclonal). Method: Rat brains were dissected and fixed overnight in 4% PFA before then being incubated in 30% sucrose (in PBS) until sunk (approx. 48hrs). A freezing microtome was used to cut 40µm horizontal slices before sections were incubated in 1% NaBH4 for 30 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% donkey serum before incubation overnight in HB6457 (1:1000, 1µg/ml) and HB6494 (1:4000 dilution). This was followed by a two hour incubation with secondary antibodies at a 1:300 dilution (polyclonal goat anti-mouse DyLight 488, Thermofisher, 35503 and goat anti-rabbit DyLight 594, Thermofisher 35561). DAPI (HB0747) was used at 1µg/ml to visualise cell nuclei. For more detail please see our IHC(IF) protocol. Images were captured using a Leica SP8 AOBS confocal laser scanning microscope attached to a Leica DM I8 inverted epifluorescence microscope. The image was captured in as a tilescan in Lightning deconvolution mode using a 63x objective (2.25x zoom), 405nm (22.3% power, PMT: 681V gain), 488nm (2.2% power, Hyd: 10% gain) and 561nm (2.1% power, Hyd: 20.8% gain) laser lines in a z-stack (0.78µm spacing). The stack was flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Figure 2. Parvalbumin and Calretinin expressing interneurons in hippocampal CA1.
Rat hippocampal CA1 stained by HB6457 for Parvalbumin (mouse monoclonal) and HB6494 for Calretinin (rabbit polyclonal) showing the different populations of interneurons in the hippocampus. Method: Rat brains were dissected and fixed overnight in 4% PFA before then being incubated in 30% sucrose (in PBS) until sunk (approx. 48hrs). A freezing microtome was used to cut 40µm horizontal slices before sections were incubated in 1% NaBH4 for 30 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% donkey serum before incubation overnight in HB6457 (1:1000, 1µg/ml) and HB6494 (1:4000 dilution). This was followed by a two hour incubation with secondary antibodies at a 1:300 dilution (polyclonal goat anti-mouse DyLight 488, Thermofisher, 35503 and goat anti-rabbit DyLight 594, Thermofisher 35561). DAPI (HB0747) was used at 1µg/ml to visualise cell nuclei. For more detail please see our IHC(IF) protocol. Images were captured using a Leica DMI6000B inverted epifluorescence microscope. The image was captured as a tilescan using a 40x objective in a z-stack (1.0µm spacing). The image was captured using DAP (95.0ms exposure), L5 (363ms exposure) and TX2 (136.1ms exposure) filters. The stack was deconvolved using Huygens professional then flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Figure 3. Parvalbumin expressing interneurons in hippocampal CA1.
Rat hippocampal CA1 stained by HB6457 for Parvalbumin (mouse monoclonal) and HB6498 for NeuN (rabbit polyclonal). Method: Rat brains were dissected and fixed overnight in 4% PFA before then being incubated in 30% sucrose (in PBS) until sunk (approx. 48hrs). A freezing microtome was used to cut 40µm horizontal slices before sections were incubated in 1% NaBH4 for 30 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% donkey serum before incubation overnight in HB6457 (1:1000, 1µg/ml) and HB6498 (1:5000 dilution). This was followed by a two hour incubation with secondary antibodies at a 1:300 dilution (polyclonal goat anti-mouse DyLight 488, Thermofisher, 35503 and goat anti-rabbit DyLight 594, Thermofisher 35561). DAPI (HB0747) was used at 1µg/ml to visualise cell nuclei. For more detail please see our IHC(IF) protocol. Images were captured using a Leica SP8 AOBS confocal laser scanning microscope attached to a Leica DM I8 inverted epifluorescence microscope. The image was captured in Lightning deconvolution mode using a 40x objective (1.28x zoom), 405nm (10.0% power, PMT: 771.1V gain), 496nm (5.6% power, Hyd: 49.2% gain) and 561nm (5.5% power, Hyd: 10% gain) laser lines in a z-stack (1.9µm spacing). The stack was flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Figure 4. Parvalbumin expressing interneurons in hippocampal CA1.
Rat hippocampal CA1 stained by HB6457 for Parvalbumin (mouse monoclonal) and HB6498 for NeuN (rabbit polyclonal). Method: Rat brains were dissected and fixed overnight in 4% PFA before then being incubated in 30% sucrose (in PBS) until sunk (approx. 48hrs). A freezing microtome was used to cut 40µm horizontal slices before sections were incubated in 1% NaBH4 for 30 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% donkey serum before incubation overnight in HB6457 (1:1000, 1µg/ml) and HB6498 (1:5000 dilution). This was followed by a two hour incubation with secondary antibodies at a 1:300 dilution (polyclonal goat anti-mouse DyLight 488, Thermofisher, 35503 and goat anti-rabbit DyLight 594, Thermofisher 35561). DAPI (HB0747) was used at 1µg/ml to visualise cell nuclei. For more detail please see our IHC(IF) protocol. Images were captured using a Leica DMI6000B inverted epifluorescence microscope. The image was captured as a tilescan using a 20x objective in a z-stack (2.4µm spacing). The image was captured using A4 (100.8ms exposure), GFP (161ms exposure) and Y3 (409.5ms exposure) filters. The stack was deconvolved using Huygens professional then flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Figure 5. Parvalbumin containing neurons stained in paraffin embedded rat cerebellum sections
HB6457 effectively stains the population of parvalbumin expressing neurons within the cerebellum of paraffin embedded horizontal rat brain sections. Method: Rat brains were dissected and fixed overnight in 4% PFA before then being incubated in 70% ethanol and embedded in paraffin. Deparaffinization was carried out following cutting of the tissue block into 5µm sections. Antigen retrieval was carried out using citrate buffer (HB8687) at 95°C for 20 minutes before sections were blocked in 1% BSA, 10% serum. HB6457 was incubated overnight (4°C) at 4µg/ml (1:250 dilution). Following washing and peroxide blocking with 0.3% H2O2, the sections were incubated in HB11345 (goat anti-mouse H&L biotin antibody) for 1 hour at room temperature (1:500 dilution). Following subsequent washes, the sections were incubated in HB5255 (Streptavidin HRP) for 30 minutes at room temperature (1:500 dilution). The reaction was developed using DAB (HB0687) and counterstained using hematoxylin (HB6189) before sections were dehydrated and mounted using DPX. For more details, please see our paraffin embedded immunohistochemistry protocol. The image was taken on a standard widefield microscope using brightfield illumination and a 5x objective.
Figure 6. Concentration response of HB6457 staining in rat hippocampus
HB6457 produces strong staining of parvalbumin positive interneurons in the hippocampus down to concentrations as low as 0.25µg/ml (1:4,000 dilution). Method: Rat brains were dissected and fixed overnight in 4% PFA before then being incubated in 30% sucrose (in PBS) until sunk (approx. 48hrs). A freezing microtome was used to cut 40µm horizontal slices before sections were incubated in 1% NaBH4 for 30 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% donkey serum before incubation overnight in HB6457 (1:500 to 1:4,000 dilutions, 0.25 - 2µg/ml). This was followed by a two hour incubation with secondary antibody at a 1:300 dilution (polyclonal goat anti-mouse DyLight 488, Thermofisher, 35503. DAPI (HB0747) was used at 1µg/ml to visualise cell nuclei. For more detail please see our IHC(IF) protocol. Images were captured using a Leica DMI6000B inverted epifluorescence microscope. Images were captured using a 10x objective in a z-stack with exposures:
1:500: DAP: 10ms, L5: 125.8ms
1:1,000: DAP: 10ms, L5: 125.8ms
1:2,000: DAP: 10ms, L5: 125.8ms
1:4,000: DAP: 10ms, L5: 106.6ms
Stacks were deconvolved using Huygens professional then flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Figure 7. Parvalbumin and Calretinin expressing interneurons in hippocampal CA1.
A section of hippocampal CA1 stained by HB6457 for Parvalbumin (mouse monoclonal) and HB6494 for Calretinin (rabbit polyclonal). Method: Rat brains were dissected and fixed overnight in 4% PFA before then being incubated in 30% sucrose (in PBS) until sunk (approx. 48hrs). A freezing microtome was used to cut 40µm horizontal slices before sections were incubated in 1% NaBH4 for 30 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% donkey serum before incubation overnight in HB6457 (1:1000, 1µg/ml) and HB6494 (1:4000 dilution). This was followed by a two hour incubation with secondary antibodies at a 1:300 dilution (polyclonal goat anti-mouse DyLight 488, Thermofisher, 35503 and goat anti-rabbit DyLight 594, Thermofisher 35561). DAPI (HB0747) was used at 1µg/ml to visualise cell nuclei. For more detail please see our IHC(IF) protocol. Images were captured using a Leica SP8 AOBS confocal laser scanning microscope attached to a Leica DM I8 inverted epifluorescence microscope. The image was captured in Lightning deconvolution mode using a 20x objective (1.28x zoom), 405nm (22.3% power, PMT: 800V gain), 488nm (2.2% power, Hyd: 15.9% gain) and 561nm (2.1% power, Hyd: 20.8% gain) laser lines in a z-stack (0.86µm spacing). The stack was flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Figure 8. Parvalbumin and Calretinin expressing neurons in rat cerebellum.
Rat cerebellum stained by HB6457 for Parvalbumin (mouse monoclonal) and HB6494 for Calretinin (rabbit polyclonal) showing the different populations of interneurons in the cerebellum. Method: Rat brains were dissected and fixed overnight in 4% PFA before then being incubated in 30% sucrose (in PBS) until sunk (approx. 48hrs). A freezing microtome was used to cut 40µm horizontal slices before sections were incubated in 1% NaBH4 for 30 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% donkey serum before incubation overnight in HB6457 (1:1000, 1µg/ml) and HB6494 (1:4000 dilution). This was followed by a two hour incubation with secondary antibodies at a 1:300 dilution (polyclonal goat anti-mouse DyLight 488, Thermofisher, 35503 and goat anti-rabbit DyLight 594, Thermofisher 35561). DAPI (HB0747) was used at 1µg/ml to visualise cell nuclei. For more detail please see our IHC(IF) protocol. Images were captured using a Leica DMI6000B inverted epifluorescence microscope. The image was captured as a tilescan using a 40x objective in a z-stack (1.0µm spacing). The image was captured using DAP (95.0ms exposure), L5 (363.0ms exposure) and TX2 (201.3ms exposure) filters. The stack was deconvolved using Huygens professional then flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Figure 9. Parvalbumin and GAT1 expressing neurons in rat cerebellum.
Rat cerebelum stained by HB6457 for Parvalbumin (mouse monoclonal) and HB7632 for GAT1 (rabbit polyclonal). Method: Rat brains were dissected and fixed overnight in 4% PFA before then being incubated in 30% sucrose (in PBS) until sunk (approx. 48hrs). A freezing microtome was used to cut 40µm horizontal slices before sections were incubated in 1% NaBH4 for 30 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% donkey serum before incubation overnight in HB6457 (1:500, 2µg/ml) and HB7632 (1:500 dilution). This was followed by a two hour incubation with secondary antibodies at a 1:300 dilution (polyclonal goat anti-mouse DyLight488, Thermofisher, 35503 and goat anti-rabbit DyLight 594, Thermofisher 35561). DAPI (HB0747) was used at 1µg/ml to visualise cell nuclei. For more detail please see our IHC(IF) protocol. Images were captured using a Leica DMI6000B inverted epifluorescence microscope. The image was captured as a tilescan using a 40x objective in a z-stack (1.5µm spacing). The image was captured using DAP (5.2ms exposure), L5 (73.5ms exposure) and TX2 (25.0ms exposure) filters. The stack was deconvolved using Huygens professional then flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Figure 10. Parvalbumin and Calretinin expressing interneurons in the rat hippocampus
Rat hippocampus stained by HB6457 for Parvalbumin (mouse monoclonal) and HB6494 for Calretinin (rabbit polyclonal) showing the different populations of interneurons in the hippocampus. Method: Rat brains were dissected and fixed overnight in 4% PFA before then being incubated in 30% sucrose (in PBS) until sunk (approx. 48hrs). A freezing microtome was used to cut 40µm horizontal slices before sections were incubated in 1% NaBH4 for 30 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% donkey serum before incubation overnight in HB6457 (1:1000, 1µg/ml) and HB6494 (1:4000 dilution). This was followed by a two hour incubation with secondary antibodies at a 1:300 dilution (polyclonal goat anti-mouse DyLight 488, Thermofisher, 35503 and goat anti-rabbit DyLight 594, Thermofisher 35561). DAPI (HB0747) was used at 1µg/ml to visualise cell nuclei. For more detail please see our IHC(IF) protocol. Images were captured using a Leica DMI6000B inverted epifluorescence microscope. The image was captured as a tilescan using a 10x objective in a z-stack (6.1µm spacing). The image was captured using DAP (205.0ms exposure), L5 (444.3ms exposure) and TX2 (231.4ms exposure) filters. The stack was deconvolved using Huygens professional then flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Figure 1. Parvalbumin and Calretinin expressing interneurons in hippocampal CA1.
A section of hippocampal CA1 stained by HB6457 for Parvalbumin (mouse monoclonal) and HB6494 for Calretinin (rabbit polyclonal). Method: Rat brains were dissected and fixed overnight in 4% PFA before then being incubated in 30% sucrose (in PBS) until sunk (approx. 48hrs). A freezing microtome was used to cut 40µm horizontal slices before sections were incubated in 1% NaBH4 for 30 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% donkey serum before incubation overnight in HB6457 (1:1000, 1µg/ml) and HB6494 (1:4000 dilution). This was followed by a two hour incubation with secondary antibodies at a 1:300 dilution (polyclonal goat anti-mouse DyLight 488, Thermofisher, 35503 and goat anti-rabbit DyLight 594, Thermofisher 35561). DAPI (HB0747) was used at 1µg/ml to visualise cell nuclei. For more detail please see our IHC(IF) protocol. Images were captured using a Leica SP8 AOBS confocal laser scanning microscope attached to a Leica DM I8 inverted epifluorescence microscope. The image was captured in as a tilescan in Lightning deconvolution mode using a 63x objective (2.25x zoom), 405nm (22.3% power, PMT: 681V gain), 488nm (2.2% power, Hyd: 10% gain) and 561nm (2.1% power, Hyd: 20.8% gain) laser lines in a z-stack (0.78µm spacing). The stack was flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Figure 2. Parvalbumin and Calretinin expressing interneurons in hippocampal CA1.
Rat hippocampal CA1 stained by HB6457 for Parvalbumin (mouse monoclonal) and HB6494 for Calretinin (rabbit polyclonal) showing the different populations of interneurons in the hippocampus. Method: Rat brains were dissected and fixed overnight in 4% PFA before then being incubated in 30% sucrose (in PBS) until sunk (approx. 48hrs). A freezing microtome was used to cut 40µm horizontal slices before sections were incubated in 1% NaBH4 for 30 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% donkey serum before incubation overnight in HB6457 (1:1000, 1µg/ml) and HB6494 (1:4000 dilution). This was followed by a two hour incubation with secondary antibodies at a 1:300 dilution (polyclonal goat anti-mouse DyLight 488, Thermofisher, 35503 and goat anti-rabbit DyLight 594, Thermofisher 35561). DAPI (HB0747) was used at 1µg/ml to visualise cell nuclei. For more detail please see our IHC(IF) protocol. Images were captured using a Leica DMI6000B inverted epifluorescence microscope. The image was captured as a tilescan using a 40x objective in a z-stack (1.0µm spacing). The image was captured using DAP (95.0ms exposure), L5 (363ms exposure) and TX2 (136.1ms exposure) filters. The stack was deconvolved using Huygens professional then flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Figure 3. Parvalbumin expressing interneurons in hippocampal CA1.
Rat hippocampal CA1 stained by HB6457 for Parvalbumin (mouse monoclonal) and HB6498 for NeuN (rabbit polyclonal). Method: Rat brains were dissected and fixed overnight in 4% PFA before then being incubated in 30% sucrose (in PBS) until sunk (approx. 48hrs). A freezing microtome was used to cut 40µm horizontal slices before sections were incubated in 1% NaBH4 for 30 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% donkey serum before incubation overnight in HB6457 (1:1000, 1µg/ml) and HB6498 (1:5000 dilution). This was followed by a two hour incubation with secondary antibodies at a 1:300 dilution (polyclonal goat anti-mouse DyLight 488, Thermofisher, 35503 and goat anti-rabbit DyLight 594, Thermofisher 35561). DAPI (HB0747) was used at 1µg/ml to visualise cell nuclei. For more detail please see our IHC(IF) protocol. Images were captured using a Leica SP8 AOBS confocal laser scanning microscope attached to a Leica DM I8 inverted epifluorescence microscope. The image was captured in Lightning deconvolution mode using a 40x objective (1.28x zoom), 405nm (10.0% power, PMT: 771.1V gain), 496nm (5.6% power, Hyd: 49.2% gain) and 561nm (5.5% power, Hyd: 10% gain) laser lines in a z-stack (1.9µm spacing). The stack was flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Figure 4. Parvalbumin expressing interneurons in hippocampal CA1.
Rat hippocampal CA1 stained by HB6457 for Parvalbumin (mouse monoclonal) and HB6498 for NeuN (rabbit polyclonal). Method: Rat brains were dissected and fixed overnight in 4% PFA before then being incubated in 30% sucrose (in PBS) until sunk (approx. 48hrs). A freezing microtome was used to cut 40µm horizontal slices before sections were incubated in 1% NaBH4 for 30 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% donkey serum before incubation overnight in HB6457 (1:1000, 1µg/ml) and HB6498 (1:5000 dilution). This was followed by a two hour incubation with secondary antibodies at a 1:300 dilution (polyclonal goat anti-mouse DyLight 488, Thermofisher, 35503 and goat anti-rabbit DyLight 594, Thermofisher 35561). DAPI (HB0747) was used at 1µg/ml to visualise cell nuclei. For more detail please see our IHC(IF) protocol. Images were captured using a Leica DMI6000B inverted epifluorescence microscope. The image was captured as a tilescan using a 20x objective in a z-stack (2.4µm spacing). The image was captured using A4 (100.8ms exposure), GFP (161ms exposure) and Y3 (409.5ms exposure) filters. The stack was deconvolved using Huygens professional then flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Figure 5. Parvalbumin containing neurons stained in paraffin embedded rat cerebellum sections
HB6457 effectively stains the population of parvalbumin expressing neurons within the cerebellum of paraffin embedded horizontal rat brain sections. Method: Rat brains were dissected and fixed overnight in 4% PFA before then being incubated in 70% ethanol and embedded in paraffin. Deparaffinization was carried out following cutting of the tissue block into 5µm sections. Antigen retrieval was carried out using citrate buffer (HB8687) at 95°C for 20 minutes before sections were blocked in 1% BSA, 10% serum. HB6457 was incubated overnight (4°C) at 4µg/ml (1:250 dilution). Following washing and peroxide blocking with 0.3% H2O2, the sections were incubated in HB11345 (goat anti-mouse H&L biotin antibody) for 1 hour at room temperature (1:500 dilution). Following subsequent washes, the sections were incubated in HB5255 (Streptavidin HRP) for 30 minutes at room temperature (1:500 dilution). The reaction was developed using DAB (HB0687) and counterstained using hematoxylin (HB6189) before sections were dehydrated and mounted using DPX. For more details, please see our paraffin embedded immunohistochemistry protocol. The image was taken on a standard widefield microscope using brightfield illumination and a 5x objective.
Figure 6. Concentration response of HB6457 staining in rat hippocampus
HB6457 produces strong staining of parvalbumin positive interneurons in the hippocampus down to concentrations as low as 0.25µg/ml (1:4,000 dilution). Method: Rat brains were dissected and fixed overnight in 4% PFA before then being incubated in 30% sucrose (in PBS) until sunk (approx. 48hrs). A freezing microtome was used to cut 40µm horizontal slices before sections were incubated in 1% NaBH4 for 30 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% donkey serum before incubation overnight in HB6457 (1:500 to 1:4,000 dilutions, 0.25 - 2µg/ml). This was followed by a two hour incubation with secondary antibody at a 1:300 dilution (polyclonal goat anti-mouse DyLight 488, Thermofisher, 35503. DAPI (HB0747) was used at 1µg/ml to visualise cell nuclei. For more detail please see our IHC(IF) protocol. Images were captured using a Leica DMI6000B inverted epifluorescence microscope. Images were captured using a 10x objective in a z-stack with exposures:
1:500: DAP: 10ms, L5: 125.8ms
1:1,000: DAP: 10ms, L5: 125.8ms
1:2,000: DAP: 10ms, L5: 125.8ms
1:4,000: DAP: 10ms, L5: 106.6ms
Stacks were deconvolved using Huygens professional then flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Figure 7. Parvalbumin and Calretinin expressing interneurons in hippocampal CA1.
A section of hippocampal CA1 stained by HB6457 for Parvalbumin (mouse monoclonal) and HB6494 for Calretinin (rabbit polyclonal). Method: Rat brains were dissected and fixed overnight in 4% PFA before then being incubated in 30% sucrose (in PBS) until sunk (approx. 48hrs). A freezing microtome was used to cut 40µm horizontal slices before sections were incubated in 1% NaBH4 for 30 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% donkey serum before incubation overnight in HB6457 (1:1000, 1µg/ml) and HB6494 (1:4000 dilution). This was followed by a two hour incubation with secondary antibodies at a 1:300 dilution (polyclonal goat anti-mouse DyLight 488, Thermofisher, 35503 and goat anti-rabbit DyLight 594, Thermofisher 35561). DAPI (HB0747) was used at 1µg/ml to visualise cell nuclei. For more detail please see our IHC(IF) protocol. Images were captured using a Leica SP8 AOBS confocal laser scanning microscope attached to a Leica DM I8 inverted epifluorescence microscope. The image was captured in Lightning deconvolution mode using a 20x objective (1.28x zoom), 405nm (22.3% power, PMT: 800V gain), 488nm (2.2% power, Hyd: 15.9% gain) and 561nm (2.1% power, Hyd: 20.8% gain) laser lines in a z-stack (0.86µm spacing). The stack was flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Figure 8. Parvalbumin and Calretinin expressing neurons in rat cerebellum.
Rat cerebellum stained by HB6457 for Parvalbumin (mouse monoclonal) and HB6494 for Calretinin (rabbit polyclonal) showing the different populations of interneurons in the cerebellum. Method: Rat brains were dissected and fixed overnight in 4% PFA before then being incubated in 30% sucrose (in PBS) until sunk (approx. 48hrs). A freezing microtome was used to cut 40µm horizontal slices before sections were incubated in 1% NaBH4 for 30 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% donkey serum before incubation overnight in HB6457 (1:1000, 1µg/ml) and HB6494 (1:4000 dilution). This was followed by a two hour incubation with secondary antibodies at a 1:300 dilution (polyclonal goat anti-mouse DyLight 488, Thermofisher, 35503 and goat anti-rabbit DyLight 594, Thermofisher 35561). DAPI (HB0747) was used at 1µg/ml to visualise cell nuclei. For more detail please see our IHC(IF) protocol. Images were captured using a Leica DMI6000B inverted epifluorescence microscope. The image was captured as a tilescan using a 40x objective in a z-stack (1.0µm spacing). The image was captured using DAP (95.0ms exposure), L5 (363.0ms exposure) and TX2 (201.3ms exposure) filters. The stack was deconvolved using Huygens professional then flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Figure 9. Parvalbumin and GAT1 expressing neurons in rat cerebellum.
Rat cerebelum stained by HB6457 for Parvalbumin (mouse monoclonal) and HB7632 for GAT1 (rabbit polyclonal). Method: Rat brains were dissected and fixed overnight in 4% PFA before then being incubated in 30% sucrose (in PBS) until sunk (approx. 48hrs). A freezing microtome was used to cut 40µm horizontal slices before sections were incubated in 1% NaBH4 for 30 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% donkey serum before incubation overnight in HB6457 (1:500, 2µg/ml) and HB7632 (1:500 dilution). This was followed by a two hour incubation with secondary antibodies at a 1:300 dilution (polyclonal goat anti-mouse DyLight488, Thermofisher, 35503 and goat anti-rabbit DyLight 594, Thermofisher 35561). DAPI (HB0747) was used at 1µg/ml to visualise cell nuclei. For more detail please see our IHC(IF) protocol. Images were captured using a Leica DMI6000B inverted epifluorescence microscope. The image was captured as a tilescan using a 40x objective in a z-stack (1.5µm spacing). The image was captured using DAP (5.2ms exposure), L5 (73.5ms exposure) and TX2 (25.0ms exposure) filters. The stack was deconvolved using Huygens professional then flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Figure 10. Parvalbumin and Calretinin expressing interneurons in the rat hippocampus
Rat hippocampus stained by HB6457 for Parvalbumin (mouse monoclonal) and HB6494 for Calretinin (rabbit polyclonal) showing the different populations of interneurons in the hippocampus. Method: Rat brains were dissected and fixed overnight in 4% PFA before then being incubated in 30% sucrose (in PBS) until sunk (approx. 48hrs). A freezing microtome was used to cut 40µm horizontal slices before sections were incubated in 1% NaBH4 for 30 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% donkey serum before incubation overnight in HB6457 (1:1000, 1µg/ml) and HB6494 (1:4000 dilution). This was followed by a two hour incubation with secondary antibodies at a 1:300 dilution (polyclonal goat anti-mouse DyLight 488, Thermofisher, 35503 and goat anti-rabbit DyLight 594, Thermofisher 35561). DAPI (HB0747) was used at 1µg/ml to visualise cell nuclei. For more detail please see our IHC(IF) protocol. Images were captured using a Leica DMI6000B inverted epifluorescence microscope. The image was captured as a tilescan using a 10x objective in a z-stack (6.1µm spacing). The image was captured using DAP (205.0ms exposure), L5 (444.3ms exposure) and TX2 (231.4ms exposure) filters. The stack was deconvolved using Huygens professional then flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Product information
Immunogen
Recombinant human parvalbumin expressed in and purified from E. coli
Clone number
3C9
Isotype
IgG1
Purification
Protein G affinity chromatography
Concentration
1mg/ml
Formulation
50% PBS, 50% glycerol + 5mM sodium azide
Predicted species reactivity
Mouse, Rat, Human, Pig, Horse, Cow
Tested species reactivity
Mouse, Rat
Tested applications
Applications
IHC-P, IHC(IF)
IHC(IF) optimal concentration
0.25µg/ml (1:4,000) as measured in free-floating paraformaldehyde fixed rat brain sections
IHC-P optimal concentration
1:250 (4µg/ml) as tested in paraffin embedded rat horizontal brain sections using streptavidin-HRP detection system.
Positive control
Parvalbumin is expressed in interneurones in a wide array of brain regions such as the cerebellum and hippocampus.
Negative control
Parvalbumin is not expressed in a range of tissues such as liver, muscle and skin in adittion to not expressing in HeLa cells.
Parvalbumin is expressed in inhibitory interneurons in various regions of the brain, including the cerebral cortex, hippocampus, and cerebellum. It is also expressed in skeletal muscle and select other tissues such as in the parathyroid gland.
Subcellular expression
Parvalbumin is expressed in the cytosol; in neurones this expression is across the whole cell body, dendritic and axonal compartments.
Processing
Following translation no processing other than having the initiator methionine removed is required for parvalbumin to reach its active conformation.
Post translational modifications
Parvalbumin is subject to phosphorylation on S2, T4 and S24 alongside acetylation on S2.
Homology (compared to human)
Compared to human parvalbumin the mouse and rat homologs show 87.3% and 91.8% identity respectively in a BLAST search. Mouse and rat parvalbumin show a 94.6% identity with 6 amino acid changes.
Similar proteins
In a BLAST search the only identified similar proteins were Oncomodulin-1 (51.4% identity, 12.1kDa) and Oncomodulin-2 (51.4% identity, 12.1kDa)
Storage & Handling
Storage instructions
-20°C
Important
This product is for RESEARCH USE ONLY and is not intended for therapeutic or diagnostic use. Not for human or veterinary use
Will my Parvalbumin antibody work against species that have not been listed on the datasheet?
A species not being listed doesn’t mean that the Parvalbumin antibody won’t work, just that we haven’t tested it. If you test one of our antibodies in a new species please let us know (positive or negative)!
What counterstains do you recommend for use in ICC and IHC with this Parvalbumin antibody?
What guarantee do you have that my Parvalbumin antibody will perform as expected?
We guarantee that your Parvalbumin antibody will work for the applications and species we list on the datasheet. If the antibody fails to perform as expected then we are happy to offer a 100% refund guarantee. For more details please see our guarantee policy.
What protocols are available for use with this Parvalbumin antibody?
We have made a comprehensive collection of protocols that we have used in our experiments to validate this Parvalbumin antibody.
What mounting media do you recommend to use with this antibody?
We recommend using one of our high performance mounting medias, supplied as either hardset or aqeous with a range of counterstains:
Reduction in parvalbumin expression not loss of the parvalbumin-expressing GABA interneuron subpopulation in genetic parvalbumin and shank mouse models of autism.
Antibody to Parvalbumin - calcium binding protein used as a marker for an inhibitory interneuron subtype. Part of the ValidAb™ range of highly validated, data-rich antibodies.
Antibody to Calretinin - calcium binding protein used as a marker for an inhibitory interneuron subtype. Part of the ValidAb™ range of highly validated, data-rich antibodies.
Antibody to Calbindin - calcium binding protein used as a marker for an inhibitory interneuron subtype. Part of the ValidAb™ range of highly validated, data-rich antibodies.
Antibody to GAT1 - GABA reuptake transporter and marker for GABAergic interneurones. Part of the ValidAb™ range of highly validated, data-rich antibodies.