Goat Anti-Mouse IgG H&L (AF488) preadsorbed ValidAb™
(HB8529)
Product overview
| Name | Goat Anti-Mouse IgG H&L (AF488) preadsorbed ValidAb™ |
| Host | Goat |
| Clonality | Polyclonal |
| Target | Mouse IgG H&L |
| Conjugate | AF488 |
| Description | Goat Anti-Mouse IgG H&L AF488 secondary antibody. Part of the ValidAb™ range of highly validated, data-rich antibodies. |
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Validation data
Figure 1. Parvalbumin and Neurofilament L and staining in rat CA1 using HB8529 Goat Anti-Mouse H&L (AF488) and HB8283 Goat Anti-Rabbit H&L (Janelia Fluor® 646)
Rat hippocampus CA1 stained for Neurofilament L (NfL, HB7266) and parvalbumin (PV, HB6457) using HB8529 goat anti-mouse IgG H&L (AF488) preadsorbed and HB8283 goat anti-rabbit IgG H&L (Janelia Fluor® 646) preadsorbed secondary antibodies. 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 15 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% goat serum before incubation overnight in HB7266 (1:4,000 dilution, 0.25µg/ml) and HB6457 (1:4,000 dilution, 0.25µg/ml). This was followed by a two-hour incubation with HB8529 goat anti-mouse IgG H&L (AF488) preadsorbed and HB8283 goat anti-rabbit IgG H&L (Janelia Fluor® 646) preadsorbed secondary antibodies at a 1:300 dilution. DAPI (HB0747) was used at 1µg/ml to visualise cell nuclei. For more detail please see our IHC(IF) protocol. The image was captured using a Leica DMI6000B fluorescence microscope with a 40x objective using DAP (8ms exposure), L5 (40ms exposure) and Y5 (157ms exposure) filters. Following capture deconvolution was carried out using Huygens Professional before the stack was flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Figure 2. Tyrosine hydroxlase and GFAP staining in rat midbrain using HB8529 Goat Anti-Mouse H&L (AF488) and HB8283 Goat Anti-Rabbit H&L (Janelia Fluor® 646)
Rat midbrain stained for Tyrosine hydroxylase (TH, HB7167) and GFAP (HB8001) using HB8529 goat anti-mouse IgG H&L (AF488) preadsorbed and HB8283 goat anti-rabbit IgG H&L (Janelia Fluor® 646) preadsorbed secondary antibodies. 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 15 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% goat serum before incubation overnight in HB8001 (1:4,000 dilution) and HB7167 (1:4,000 dilution). This was followed by a two-hour incubation with HB8529 goat anti-mouse IgG H&L (AF488) preadsorbed and HB8283 goat anti-rabbit IgG H&L (Janelia Fluor® 646) preadsorbed secondary antibodies at a 1:300 dilution. DAPI (HB0747) was used at 1µg/ml to visualise cell nuclei. For more detail please see our IHC(IF) protocol. The image was captured using a Leica DMI6000B fluorescence microscope with a 20x objective using DAP (20ms exposure), L5 (61ms exposure) and RHO (250ms exposure) filters. Following capture deconvolution was carried out using Huygens Professional before the stack was flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Figure 3. Parvalbumin and Neurofilament L and staining in rat cerebellum using HB8529 Goat Anti-Mouse H&L (AF488) and HB8283 Goat Anti-Rabbit H&L (Janelia Fluor® 646)
Rat cerebellum stained for Neurofilament L (NfL, HB7266) and parvalbumin (PV, HB6457) using HB8529 goat anti-mouse IgG H&L (AF488) preadsorbed and HB8283 goat anti-rabbit IgG H&L (Janelia Fluor® 646) preadsorbed secondary antibodies. 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 15 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% goat serum before incubation overnight in HB7266 (1:4,000 dilution, 0.25µg/ml) and HB6457 (1:4,000 dilution, 0.25µg/ml). This was followed by a two-hour incubation with HB8529 goat anti-mouse IgG H&L (AF488) preadsorbed and HB8283 goat anti-rabbit IgG H&L (Janelia Fluor® 646) preadsorbed secondary antibodies at a 1:300 dilution. DAPI (HB0747) was used at 1µg/ml to visualise cell nuclei. For more detail please see our IHC(IF) protocol. The image was captured using a Leica DMI6000B fluorescence microscope with a 20x objective using DAP (30ms exposure), L5 (150ms exposure) and Y5 (250ms exposure) filters. Following capture deconvolution was carried out using Huygens Professional before the stack was flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Figure 4. Vimentin staining in HEK293T cells using HB8529 Goat Anti-Mouse H&L (AF488).
HEK293T cells stained with for Vimentin (HB6497) using HB8529 goat anti-mouse IgG H&L (AF488) preadsorbed as the secondary antibody. Method: HEK293 cells were cultured following established protocols (Lee et al., 2019. PLoS ONE, 14(5):e0213116) before being fixed with 4% PFA. Cells were permeabilised with 0.1% Triton X-100 followed by blocking in 1% BSA, 300mM glycine. HB6497 (mouse monoclonal anti-vimentin) was incubated overnight (4°C) at a 1:2,000 dilution (0.5µg/ml) followed by a one hour incubation with HB8529 goat anti-mouse IgG H&L (AF488) preadsorbed secondary antibody (1:300 dilution). DAPI (HB0747) was used at 1µg/ml to visualise cell nuclei. For more detail please see our ICC protocol. Images were captured as a tilescan using a Leica DMI6000B inverted epifluorescence microscope with a 40x objective and DAP (6ms exposure) / L5 (26ms exposure) filters. Images were captured as a z-stack (0.30µm spacing) before being deconvolved using Huygens Essential (Scientific Volume Imagine) and flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Figure 5. Neurofilament L and Parvalbumin staining in rat CA1 using HB8529 Goat Anti-Mouse H&L (AF488) and HB8283 Goat Anti-Rabbit H&L (Janelia Fluor® 646)
Rat hippocampus CA1 stained for Neurofilament L (NfL, HB7266) and parvalbumin (PV, HB6457) using HB8529 goat anti-mouse IgG H&L (AF488) preadsorbed and HB8283 goat anti-rabbit IgG H&L (Janelia Fluor® 646) preadsorbed secondary antibodies. 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 15 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% goat serum before incubation overnight in HB7266 (1:4,000 dilution, 0.25µg/ml) and HB6457 (1:4,000 dilution, 0.25µg/ml). This was followed by a two-hour incubation with HB8529 goat anti-mouse IgG H&L (AF488) preadsorbed and HB8283 goat anti-rabbit IgG H&L (Janelia Fluor® 646) preadsorbed secondary antibodies at a 1:300 dilution. DAPI (HB0747) was used at 1µg/ml to visualise cell nuclei. For more detail please see our IHC(IF) protocol. The image was captured using a Leica DMI6000B fluorescence microscope with a 40x objective using DAP (8ms exposure), L5 (40ms exposure) and Y5 (157ms exposure) filters. Following capture, deconvolution was carried out using Huygens Professional before the stack was flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Figure 6. Tyrosine hydroxlase staining in rat caudate putamen using HB8529 Goat Anti-Mouse H&L (AF488) secondary antibody
Rat caudate putamen (CPu) stained for Tyrosine hydroxylase (TH) with HB7167) using HB8529 goat anti-mouse IgG H&L (AF488) preadsorbed as the secondary antibody. 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 15 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% goat serum before incubation overnight in HB7167 (1:4,000 dilution). This was followed by a two-hour incubation with HB8529 goat anti-mouse IgG H&L (AF488) preadsorbed secondary antibody at a 1:300 dilution. DAPI (HB0747) was used at 1µg/ml to visualise cell nuclei. For more detail please see our IHC(IF) protocol. The image was captured using a Leica DMI6000B fluorescence microscope with a 40x objective using DAP (5ms exposure), L5 (27ms exposure) and Y5 (59ms exposure) filters. Following capture deconvolution was carried out using Huygens Professional before the stack was flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Figure 7. Tyrosine hydroxlase staining in rat midbrain using HB8529 Goat Anti-Mouse H&L (AF488) secondary antibody
Rat midbrain stained for Tyrosine hydroxylase (TH) with HB7167) using HB8529 goat anti-mouse IgG H&L (AF488) preadsorbed as the secondary antibody. 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 15 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% goat serum before incubation overnight in HB7167 (1:4,000 dilution). This was followed by a two-hour incubation with HB8529 goat anti-mouse IgG H&L (AF488) preadsorbed secondary antibody at a 1:300 dilution. DAPI (HB0747) was used at 1µg/ml to visualise cell nuclei. For more detail please see our IHC(IF) protocol. The image was captured using a Leica DMI6000B fluorescence microscope with a 40x objective using DAP (5ms exposure), L5 (27ms exposure) and Y5 (30ms exposure) filters. Following capture deconvolution was carried out using Huygens Professional before the stack was flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Figure 8. Tyrosine hydroxlase and GFAP staining in rat midbrain using HB8529 Goat Anti-Mouse H&L (AF488) and HB8283 Goat Anti-Rabbit H&L (Janelia Fluor® 646)
Rat midbrain stained for Tyrosine hydroxylase (TH, HB7167) and GFAP (HB8001) using HB8529 goat anti-mouse IgG H&L (AF488) preadsorbed and HB8283 goat anti-rabbit IgG H&L (Janelia Fluor® 646) preadsorbed secondary antibodies. 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 15 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% goat serum before incubation overnight in HB8001 (1:4,000 dilution) and HB7167 (1:4,000 dilution). This was followed by a two-hour incubation with HB8529 goat anti-mouse IgG H&L (AF488) preadsorbed and HB8283 goat anti-rabbit IgG H&L (Janelia Fluor® 646) preadsorbed secondary antibodies at a 1:300 dilution. 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 (2.25x zoom), 405nm (15.5% power, PMT: 723.5V gain), 488nm (1.5% power, Hyd: 10% gain) and 633nm (0.5% power, Hyd: 10% gain) laser lines in a z-stack (0.54µm spacing). The stack was flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Figure 1. Parvalbumin and Neurofilament L and staining in rat CA1 using HB8529 Goat Anti-Mouse H&L (AF488) and HB8283 Goat Anti-Rabbit H&L (Janelia Fluor® 646)
Rat hippocampus CA1 stained for Neurofilament L (NfL, HB7266) and parvalbumin (PV, HB6457) using HB8529 goat anti-mouse IgG H&L (AF488) preadsorbed and HB8283 goat anti-rabbit IgG H&L (Janelia Fluor® 646) preadsorbed secondary antibodies. 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 15 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% goat serum before incubation overnight in HB7266 (1:4,000 dilution, 0.25µg/ml) and HB6457 (1:4,000 dilution, 0.25µg/ml). This was followed by a two-hour incubation with HB8529 goat anti-mouse IgG H&L (AF488) preadsorbed and HB8283 goat anti-rabbit IgG H&L (Janelia Fluor® 646) preadsorbed secondary antibodies at a 1:300 dilution. DAPI (HB0747) was used at 1µg/ml to visualise cell nuclei. For more detail please see our IHC(IF) protocol. The image was captured using a Leica DMI6000B fluorescence microscope with a 40x objective using DAP (8ms exposure), L5 (40ms exposure) and Y5 (157ms exposure) filters. Following capture deconvolution was carried out using Huygens Professional before the stack was flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Figure 2. Tyrosine hydroxlase and GFAP staining in rat midbrain using HB8529 Goat Anti-Mouse H&L (AF488) and HB8283 Goat Anti-Rabbit H&L (Janelia Fluor® 646)
Rat midbrain stained for Tyrosine hydroxylase (TH, HB7167) and GFAP (HB8001) using HB8529 goat anti-mouse IgG H&L (AF488) preadsorbed and HB8283 goat anti-rabbit IgG H&L (Janelia Fluor® 646) preadsorbed secondary antibodies. 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 15 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% goat serum before incubation overnight in HB8001 (1:4,000 dilution) and HB7167 (1:4,000 dilution). This was followed by a two-hour incubation with HB8529 goat anti-mouse IgG H&L (AF488) preadsorbed and HB8283 goat anti-rabbit IgG H&L (Janelia Fluor® 646) preadsorbed secondary antibodies at a 1:300 dilution. DAPI (HB0747) was used at 1µg/ml to visualise cell nuclei. For more detail please see our IHC(IF) protocol. The image was captured using a Leica DMI6000B fluorescence microscope with a 20x objective using DAP (20ms exposure), L5 (61ms exposure) and RHO (250ms exposure) filters. Following capture deconvolution was carried out using Huygens Professional before the stack was flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Figure 3. Parvalbumin and Neurofilament L and staining in rat cerebellum using HB8529 Goat Anti-Mouse H&L (AF488) and HB8283 Goat Anti-Rabbit H&L (Janelia Fluor® 646)
Rat cerebellum stained for Neurofilament L (NfL, HB7266) and parvalbumin (PV, HB6457) using HB8529 goat anti-mouse IgG H&L (AF488) preadsorbed and HB8283 goat anti-rabbit IgG H&L (Janelia Fluor® 646) preadsorbed secondary antibodies. 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 15 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% goat serum before incubation overnight in HB7266 (1:4,000 dilution, 0.25µg/ml) and HB6457 (1:4,000 dilution, 0.25µg/ml). This was followed by a two-hour incubation with HB8529 goat anti-mouse IgG H&L (AF488) preadsorbed and HB8283 goat anti-rabbit IgG H&L (Janelia Fluor® 646) preadsorbed secondary antibodies at a 1:300 dilution. DAPI (HB0747) was used at 1µg/ml to visualise cell nuclei. For more detail please see our IHC(IF) protocol. The image was captured using a Leica DMI6000B fluorescence microscope with a 20x objective using DAP (30ms exposure), L5 (150ms exposure) and Y5 (250ms exposure) filters. Following capture deconvolution was carried out using Huygens Professional before the stack was flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Figure 4. Vimentin staining in HEK293T cells using HB8529 Goat Anti-Mouse H&L (AF488).
HEK293T cells stained with for Vimentin (HB6497) using HB8529 goat anti-mouse IgG H&L (AF488) preadsorbed as the secondary antibody. Method: HEK293 cells were cultured following established protocols (Lee et al., 2019. PLoS ONE, 14(5):e0213116) before being fixed with 4% PFA. Cells were permeabilised with 0.1% Triton X-100 followed by blocking in 1% BSA, 300mM glycine. HB6497 (mouse monoclonal anti-vimentin) was incubated overnight (4°C) at a 1:2,000 dilution (0.5µg/ml) followed by a one hour incubation with HB8529 goat anti-mouse IgG H&L (AF488) preadsorbed secondary antibody (1:300 dilution). DAPI (HB0747) was used at 1µg/ml to visualise cell nuclei. For more detail please see our ICC protocol. Images were captured as a tilescan using a Leica DMI6000B inverted epifluorescence microscope with a 40x objective and DAP (6ms exposure) / L5 (26ms exposure) filters. Images were captured as a z-stack (0.30µm spacing) before being deconvolved using Huygens Essential (Scientific Volume Imagine) and flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Figure 5. Neurofilament L and Parvalbumin staining in rat CA1 using HB8529 Goat Anti-Mouse H&L (AF488) and HB8283 Goat Anti-Rabbit H&L (Janelia Fluor® 646)
Rat hippocampus CA1 stained for Neurofilament L (NfL, HB7266) and parvalbumin (PV, HB6457) using HB8529 goat anti-mouse IgG H&L (AF488) preadsorbed and HB8283 goat anti-rabbit IgG H&L (Janelia Fluor® 646) preadsorbed secondary antibodies. 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 15 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% goat serum before incubation overnight in HB7266 (1:4,000 dilution, 0.25µg/ml) and HB6457 (1:4,000 dilution, 0.25µg/ml). This was followed by a two-hour incubation with HB8529 goat anti-mouse IgG H&L (AF488) preadsorbed and HB8283 goat anti-rabbit IgG H&L (Janelia Fluor® 646) preadsorbed secondary antibodies at a 1:300 dilution. DAPI (HB0747) was used at 1µg/ml to visualise cell nuclei. For more detail please see our IHC(IF) protocol. The image was captured using a Leica DMI6000B fluorescence microscope with a 40x objective using DAP (8ms exposure), L5 (40ms exposure) and Y5 (157ms exposure) filters. Following capture, deconvolution was carried out using Huygens Professional before the stack was flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Figure 6. Tyrosine hydroxlase staining in rat caudate putamen using HB8529 Goat Anti-Mouse H&L (AF488) secondary antibody
Rat caudate putamen (CPu) stained for Tyrosine hydroxylase (TH) with HB7167) using HB8529 goat anti-mouse IgG H&L (AF488) preadsorbed as the secondary antibody. 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 15 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% goat serum before incubation overnight in HB7167 (1:4,000 dilution). This was followed by a two-hour incubation with HB8529 goat anti-mouse IgG H&L (AF488) preadsorbed secondary antibody at a 1:300 dilution. DAPI (HB0747) was used at 1µg/ml to visualise cell nuclei. For more detail please see our IHC(IF) protocol. The image was captured using a Leica DMI6000B fluorescence microscope with a 40x objective using DAP (5ms exposure), L5 (27ms exposure) and Y5 (59ms exposure) filters. Following capture deconvolution was carried out using Huygens Professional before the stack was flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Figure 7. Tyrosine hydroxlase staining in rat midbrain using HB8529 Goat Anti-Mouse H&L (AF488) secondary antibody
Rat midbrain stained for Tyrosine hydroxylase (TH) with HB7167) using HB8529 goat anti-mouse IgG H&L (AF488) preadsorbed as the secondary antibody. 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 15 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% goat serum before incubation overnight in HB7167 (1:4,000 dilution). This was followed by a two-hour incubation with HB8529 goat anti-mouse IgG H&L (AF488) preadsorbed secondary antibody at a 1:300 dilution. DAPI (HB0747) was used at 1µg/ml to visualise cell nuclei. For more detail please see our IHC(IF) protocol. The image was captured using a Leica DMI6000B fluorescence microscope with a 40x objective using DAP (5ms exposure), L5 (27ms exposure) and Y5 (30ms exposure) filters. Following capture deconvolution was carried out using Huygens Professional before the stack was flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Figure 8. Tyrosine hydroxlase and GFAP staining in rat midbrain using HB8529 Goat Anti-Mouse H&L (AF488) and HB8283 Goat Anti-Rabbit H&L (Janelia Fluor® 646)
Rat midbrain stained for Tyrosine hydroxylase (TH, HB7167) and GFAP (HB8001) using HB8529 goat anti-mouse IgG H&L (AF488) preadsorbed and HB8283 goat anti-rabbit IgG H&L (Janelia Fluor® 646) preadsorbed secondary antibodies. 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 15 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% goat serum before incubation overnight in HB8001 (1:4,000 dilution) and HB7167 (1:4,000 dilution). This was followed by a two-hour incubation with HB8529 goat anti-mouse IgG H&L (AF488) preadsorbed and HB8283 goat anti-rabbit IgG H&L (Janelia Fluor® 646) preadsorbed secondary antibodies at a 1:300 dilution. 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 (2.25x zoom), 405nm (15.5% power, PMT: 723.5V gain), 488nm (1.5% power, Hyd: 10% gain) and 633nm (0.5% power, Hyd: 10% gain) laser lines in a z-stack (0.54µm spacing). The stack was flattened using a maximum Z projection in ImageJ (Schindelin et al., 2012. Nat Methods, 9(7), 676–682).
Product information
| Immunogen | Purified mouse IgG |
| Purification | Immunogen affinity chromatography. Pre-adsorbed with bovine, horse, human, pig and rabbit serum proteins |
| Concentration | 1mg/ml |
| Formulation | 20% glycerol in PBS with 0.05% sodium azide and 1% recombinant albumin |
Tested applications
| Applications | ELISA, FACS and flow cytometry, ICC, IHC(IF) |
| IHC(IF) optimal concentration | 1:300 to 1:2,000 dilution (0.5 - 3.3µg/ml). Optimise dependent upon assay. A good starting point is 1:500 (2µg/ml). |
| ICC optimal concentration | 1:300 to 1:2,000 dilution (0.5 - 3.3µg/ml). Optimise dependent upon assay. A good starting point is 1:500 (2µg/ml). |
| Negative control | While this antibody has been cross-adsorbed to reduce non-specific binding it is still often worthwhile to conduct a control experiment where the primary antibody is omitted to give confidence that the staining pattern observed is specific. |
Optical Data
| Max excitation wavelength | 490 nm |
| Max emission wavelength | 525 nm |
| Emission color | Green |
| Closest laser lines | 488 nm |
| Spectrally similar dyes | Alexa Fluor® 488, Atto 488, DyLight® 488, FITC |
| Fluorescence spectra |  |
Storage & Handling
| Storage instructions | +4°C |
| Important | This product is for RESEARCH USE ONLY and is not intended for therapeutic or diagnostic use. Not for human or veterinary use |
Technical guides
Western Blot Protocol (1 MB) Immunocytochemistry Protocol (1.2 MB) Immunohistochemistry Protocol (1.6 MB) References for Goat Anti-Mouse IgG H&L (AF488) preadsorbed ValidAb™
References are publications that support the biological activity of the product
-
Human Neutrophils Present Mild Activation by Zika Virus But Reduce the Infection of Susceptible Cells.
Aggio JB et al (2022) Frontiers in immunology 13 : 784443 -
The Role of Wnt/β-Catenin Pathway Mediators in Aortic Valve Stenosis.
Khan K et al (2020) Frontiers in cell and developmental biology 8 : 862
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