HEK293T cells were lysed in 1× lysis buffer on ice for 30 min and centrifuged at 12,000 rpm for 15 min. Cell lysates were separated by electrophoresis on 4 to 20% SDS–polyacrylamide gel electrophoresis gels and then transferred onto nitrocellulose membranes. The membranes were blocked with 5% milk in TBST (Tris-buffered saline, 0.1% Tween® 20) for 1 hour and incubated overnight at 4°C with the corresponding primary antibodies: rabbit anti-FLAG and anti–β-tubulin (Sigma-Aldrich), rabbit anti-H3K9me1 (Abcam), and rabbit anti-H2AK119ub (Cell Signaling Technology). Next, the experiment was followed by incubation with the horseradish peroxidase–labeled Goat Anti-Rabbit immunoglobulin G (Beyotime) for 1 hour at room temperature 20° to 25°C. Quantitation of immunoblots was then performed via densitometric analysis using the ImageJ software.
Goat anti rabbit immunoglobulin g
Manufactured by Beyotime
Goat Anti-Rabbit immunoglobulin G is a secondary antibody used in various immunoassays and other laboratory techniques. It is a polyclonal antibody produced by immunizing goats with rabbit immunoglobulin G and purifying the resulting antibodies. This product can be used to detect and quantify rabbit primary antibodies in samples.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using goat anti rabbit immunoglobulin g
1
Western Blot Analysis of Histone Modifications
2
Immunocytochemistry Analysis of HSF1 and Notch1
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Cultured NRVMs were washed three times with PBS, fixed in 4% paraformaldehyde for 30 min, permeabilized with 0.3% Triton X-100 for 1 h, and then blocked for 1 h in PBS with goat serum. Next, the cells were incubated with primary antibodies against HSF1 (1 : 100; ThermoFisher Scientific) or Notch1 (1 : 100; ThermoFisher Scientific) overnight at 4°C. Afterward, the sections were incubated with goat anti-rabbit immunoglobulin G (heavy + light chains) superclonal secondary antibody or goat anti-mouse immunoglobulin G secondary antibody (Beyotime) for 1 h. Cell nuclei were counterstained with DAPI (Beyotime). The sections were imaged using a laser scanning confocal microscope (Nikon, Tokyo, Japan) and analyzed with ImageJ.
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