Anti-Parvalbumin antibody ValidAbTM

(HB6457)
Technical documents: SDS Datasheet

Product overview

Name Anti-Parvalbumin antibody ValidAbTM
Host Mouse
Clonality Monoclonal
Target Parvalbumin
Description

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.

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Validation data

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. 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 6. 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 7. 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 8. 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 9. 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(IF)
IHC(IF) optimal concentration

0.25µg/ml (1:4,000) as measured in free-floating paraformaldehyde fixed rat brain sections

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.

Open data link

Please follow this link to OSF

Target information

Other names

Parvalbumin alpha, PV, PVALB, D22S749

UniProt ID P20472
Structure image  Chemical Structure
Gene name PVALB
NCBI full gene name Parvalbumin
Entrez gene ID

5816

Amino acids

110 (12.1kDa)

Isoforms

Parvalbumin has only one described isoform

Expression

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

FAQs

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?

We recommend using either DAPI, Hoechst 33342 or Propidium Iodide to label cell nuclei.

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?
Any other questions?

For any other questions about our antibody products please see our technical FAQs for antibodies

References for Anti-Parvalbumin antibody ValidAbTM

References are publications that support the biological activity of the product
  • The Role of Parvalbumin Interneurons in Neurotransmitter Balance and Neurological Disease.

    Nahar L et al (2021) Frontiers in psychiatry 12 : 679960
  • Parvalbumin interneuron vulnerability and brain disorders.

    Ruden JB et al (2021) Neuropsychopharmacology : official publication of the American College of Neuropsychopharmacology 46 : 279-287
  • Reduction in parvalbumin expression not loss of the parvalbumin-expressing GABA interneuron subpopulation in genetic parvalbumin and shank mouse models of autism.

    Filice F et al (2016) Molecular brain 9 : 10
  • Parvalbumin-positive interneurons of the prefrontal cortex support working memory and cognitive flexibility.

    Murray AJ et al (2015) Scientific reports 5 : 16778
  • Role of the calcium-binding protein parvalbumin in short-term synaptic plasticity.

    Caillard O et al (2000) Proceedings of the National Academy of Sciences of the United States of America 97 : 13372-7