Cell lysates were boiled in sample buffer containing 100 mM DTT for 5 min, resolved by 4 to 12% gradient SDS-PAGE, and transferred to nitrocellulose. Primary antibodies were used at 1:1000 dilution in tris-buffered saline with Tween 20 (0.1% Tween 20) plus 5% bovine serum albumin, with the exception of anti-HSP90, which was used at 1:2000 dilution. Secondary antibodies [goat anti-rabbit immunoglobulin G (IgG) horseradish peroxidase (HRP), goat anti-mouse IgG HRP (both from Bio-Rad), and goat anti-guinea pig IgG HRP (Jackson ImmunoResearch)] were used at 1:5000 dilution. Imaging was performed after incubation with the Clarity Western ECL Substrate (Bio-Rad no. 1705061) according to the manufacturer’s instructions.
Goat anti guinea pig igg hrp
Manufactured by Jackson ImmunoResearch
Goat anti-guinea pig IgG HRP is a secondary antibody that binds to guinea pig immunoglobulin G (IgG) and is conjugated with horseradish peroxidase (HRP). The HRP enzyme can be used to detect and visualize the bound primary antibody in various immunoassay techniques.
Automatically generated - may contain errors
2 protocols using goat anti guinea pig igg hrp
1
Western Blot Protein Analysis Procedure
2
PRDM9 and HELLS Expression Analysis in Mouse Testes
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For PRDM9 and HELLS expression analysis in testes from 22 dpp Hells CTRL and Hells cKO mice, nuclear fractions (40 µg) were analyzed by western blotting with affinity-purified rabbit anti-PRDM9 (1/2,000) (Grey et al., 2017 (link)) and rabbit anti-HELLS (NB100-278, Novus) (1/2,000) antibodies and Guinea-pig serum raised against the mouse SYCP3 residues 24–44 (1/2,000). Secondary antibodies were goat anti-rabbit IgG-HRP (1/10,000) (1858415, Pierce) and goat anti-Guinea-pig IgG-HRP (1/3,000) (706-035-148, Jackson Immuno Research).
For PRDM9 and HELLS expression during mouse spermatogenesis, 50 μg of whole cell extracts were analyzed by western blotting with affinity-purified rabbit anti-PRDM9 (Grey et al., 2017 (link)), mouse anti-HELLS (SC-46665, Santa Cruz) and rat anti-tubulin (ab6161, Abcam) antibodies.
For PRDM9 and HELLS expression during mouse spermatogenesis, 50 μg of whole cell extracts were analyzed by western blotting with affinity-purified rabbit anti-PRDM9 (Grey et al., 2017 (link)), mouse anti-HELLS (SC-46665, Santa Cruz) and rat anti-tubulin (ab6161, Abcam) antibodies.
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