Total protein was extracted from the cells as described above. The proteins (50 µg) were separated using 10% SDS-PAGE gels and transferred to nitrocellulose membranes. The membranes were blocked with 5% non-fat milk in TBST for 1 h at 37°C and incubated with primary antibodies against Bax (sc-6236; 1:10,000), PI3K (sc-293172; 1:2,000), phosphorylated (p)-Akt (sc-7985-R; 1:1,000), Akt (sc-135829; 1:1,000), p-mTOR (sc-293133; 1:1,000), mTOR (sc-1549; 1:1,000), and GADPH (sc-47724; 1:1,000; all Santa Cruz Biotechnology, Inc., Dallas, TX, USA) overnight at 4°C. The membranes were subsequently incubated with goat anti-rabbit IgG-horseradish peroxidase secondary antibodies (sc-2004; 1:5,000; Santa Cruz Biotechnology, Inc.) at 37°C for 1 h and developed using an enhanced chemiluminescent detection system (Beyotime Institute of Biotechnology). The protein bands were scanned using a Fujifilm LAS-3000 Imaging system (Fujifilm Corporation, Tokyo, Japan) and analyzed with Image Lab 3.0 (Bio-Rad Laboratories, Inc.).
Goat anti rabbit igg horseradish peroxidase secondary antibodies
Manufactured by Santa Cruz Biotechnology
Goat anti-rabbit IgG-horseradish peroxidase secondary antibodies are a type of laboratory reagent used in various immunoassay techniques. They are composed of goat-derived antibodies that specifically recognize and bind to rabbit immunoglobulin G (IgG) molecules. The antibodies are conjugated with the enzyme horseradish peroxidase, which can catalyze a colorimetric or chemiluminescent reaction for detection and quantification purposes.
Automatically generated - may contain errors
Lab products found in correlation
Las 3000 imaging system, by Fujifilm (1 mentions)
Image lab 3, by Bio-Rad (1 mentions)
Enhanced chemiluminescent detection system, by Beyotime (1 mentions)
Gadph, by Santa Cruz Biotechnology (1 mentions)
Sc 293133, by Santa Cruz Biotechnology (1 mentions)
Sc 2004, by Santa Cruz Biotechnology (1 mentions)
2 protocols using goat anti rabbit igg horseradish peroxidase secondary antibodies
1
Western Blot Analysis of Apoptosis and Survival Signaling
2
Quantitative Analysis of Retinal Proteins
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The Retinas were collected and homogenized in buffer containing 0.23 mol sucrose, 2 mmol EDTA, 5 mmol Tris–HCl (pH 7.5), and 0.1 mmol phenylmethylsulfonyl fluoride. After centrifugation, aliquot extracts containing equal amounts of protein (20 μg) were electrophoresed, transferred, and probed a polyclonal rabbit anti-cone arrestin (1:1000, sc-67212, Santa Cruz Biotechnology, CA) or polyclonal rabbit anti-rhodopsin (1:1000, sc-15382, Santa Cruz Biotechnology, CA). The membrane was washed thoroughly and then incubated with goat anti-rabbit IgG horseradish peroxidase secondary antibodies (1:3000, sc-2004, Santa Cruz Biotechnology, CA). Visualization of immunoreactive bands was performed using the Chemiluminescence Fluorescence Imaging System (Super Signal ECL kit; West Pico; Pierce, Rockford, IL, USA).
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