Total proteins were extracted from the samples and quantified using ice-cold radioimmunoprecipitation assay lysis buffer and the bicinchoninic acid protein assay kit (Pierce, Rockford, IL). Equal quantities of protein were separated on 10% sodium dodecyl sulfate polyacrylamide gels and transferred onto polyvinylidene difluoride membranes (Amersham Pharmacia Biotech), which were then blocked at room temperature for 1 hour in Tris-buffered saline with 0.2% Tween 20 containing 5% skim milk and probed with primary antibodies overnight at 4°C. Secondary antibodies included horseradish peroxidase label and were incubated for 1 hour at room temperature. The diluted concentrations of the primary antibodies were as follows: anti-human RYBP, GAPDH, EGFR, and phosphorylated-EGFR antibodies were from Sigma (St. Louis, MO); anti-human AKT, phospho-AKT, ERK, and phospho-ERK were from CST; anti-human E-cadherin and vimentin were from Santa Cruz Biotechnology.
Anti human rybp
Manufactured by Merck Group
Anti-human RYBP is a laboratory reagent used for the detection and quantification of RYBP protein in human samples. It is a monoclonal antibody that specifically binds to the RYBP protein, allowing researchers to measure its expression levels in cells or tissues.
Automatically generated - may contain errors
Lab products found in correlation
Anti human e cadherin, by Santa Cruz Biotechnology (1 mentions)
Polyvinylidene difluoride membrane, by Cytiva (1 mentions)
Gapdh, by Merck Group (1 mentions)
Vimentin, by Santa Cruz Biotechnology (1 mentions)
Goat anti mouse igg h l, by Bio-Rad (1 mentions)
Goat anti rabbit igg h l, by Bio-Rad (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Vimentin, by Merck Group (1 mentions)
β actin, by Merck Group (1 mentions)
2 protocols using anti human rybp
1
Western Blot Protocol for Protein Quantification
2
Molecular Markers for Cancer Progression
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All chemicals and solvents were of the highest analytical grade available. Cell culture supplies and media, fetal bovine serum (FBS), phosphate-buffered saline (PBS), and penicillin-streptomycin were obtained from Invitrogen (Carlsbad, CA). The anti-human p53, Bax, and poly (ADP-ribose) polymerase (PARP) antibodies were from Santa Cruz Biotechnology Inc. (Dallas, TX). The anti-human E-cadherin antibody was from BD Biosciences (San Jose, CA). The anti-human RYBP, vimentin, and β-actin antibodies were from Sigma (St. Louis, MO). Goat anti-mouse IgG (H+L) and goat anti-rabbit IgG (H+L) were obtained from Bio-Rad (Hercules, CA). The preparation of the Myc-RYBP expression vector was described previously. The RYBP siRNA pool and control siRNA pool were obtained from Dharmacon.
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