Anti-HPV16E6 antibodies 1E-6F4 (E6 N-ter) and 2E-3F8 (E6 C-ter) were obtained from Euromedex. Anti-p53 (DO-1) and anti-pRb (4H1) antibodies were obtained from Cell Signaling. Anti-β-actin (AC-15) antibody was obtained from Sigma-Aldrich. Anti-HPV16E7 (NM2) antibody was obtained from Santa Cruz Biotechnology. HRP-conjugated goat anti-mouse secondary antibody was purchased from BD Pharmingen. Primary and secondary antibodies have been diluted respectively to 1:1000 and 1:5000 in blocking buffer (except for β-actin at 1:20,000 and 1:30,000).
Anti prb 4h1
Manufactured by Cell Signaling Technology
Sourced in United States
Anti-pRb (4H1) is an antibody product offered by Cell Signaling Technology. It is used for the detection of phosphorylated retinoblastoma protein (pRb), which plays a crucial role in cell cycle regulation.
Automatically generated - may contain errors
Lab products found in correlation
Anti flag m2, by Merck Group (2 mentions)
Nanodrop spectrophotometer, by Thermo Fisher Scientific (2 mentions)
Anti p53 do 1, by Cell Signaling Technology (1 mentions)
Anti β actin antibody ac 15, by Merck Group (1 mentions)
Ab16128, by Abcam (1 mentions)
Anti ezh2 ac22, by Cell Signaling Technology (1 mentions)
Anti p65, by Cell Signaling Technology (1 mentions)
Anti e2f1 kh95, by Santa Cruz Biotechnology (1 mentions)
Anti β actin c4, by MP Biomedicals (1 mentions)
Anti phosphor iκbα, by Cell Signaling Technology (1 mentions)
Anti total iκbα, by Cell Signaling Technology (1 mentions)
Anti α tubulin b 5 1 2, by Merck Group (1 mentions)
Ecl prime western blotting detection reagent, by GE Healthcare (1 mentions)
Chemidoc xr, by Bio-Rad (1 mentions)
Benzonase, by Merck Group (1 mentions)
Anti p53 do 1, by Santa Cruz Biotechnology (1 mentions)
Goat anti mouse igg h l horseradish peroxidase hrp, by Jackson ImmunoResearch (1 mentions)
4 protocols using anti prb 4h1
1
Immunodetection of HPV16 Proteins
2
Comprehensive Immunoblotting Analysis of EBV Proteins
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The following antibodies were used: anti-DOK1 (ab8112, Abcam), anti-E2F1 (KH-95; Santa Cruz Biotechnology), anti- β-Actin C4 (MP Biomedicals), anti-LMP1 (S12), anti- phosphor IκBα (#9246, Cell Signaling Technology), anti-total IκBα (#9242, Cell Signaling Technology), mouse IgG, rabbit IgG (Santa Cruz Biotechnology), anti-p65 (#3034, Cell Signaling Technology), anti-H3K4me3, and anti-H3K27me3 (Epigentek), anti-EZH2 (AC22; Cell Signaling Technology), anti-pRB (4H1, Cell Signaling Technology), anti-DNMT1 (60B1220, Abnova), anti-EBNA1 (1EB12, Santa Cruz Biotechnology), anti-EBNA2 (Novocastra), anti-EBNA3A (Exalpha), anti-EBNA3C (ab16128, Abcam). Immunoblotting was performed as described previously [29] (link).
3
Protein Extraction and Western Blot Analysis
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The total cell extract was prepared from the recovered cells by incubation in RIPA buffer (20 mM Tris-HCl (pH 8.0), 150 mM NaCl, 5 mM MgCl2, 1 mM EDTA, 1 mM dithiothreitol, 1% Nonidet P-40, and complete protease inhibitor cocktail (Roche Diagnostic)) at 4°C for 10 min, followed by centrifugation at 21,000× g at 4 °C for 10 min. Proteins in the extract were separated on a 4–15% polyacrylamide gel using SDS-PAGE, transferred to a PVDF membrane, and detected by the ECL prime western blotting detection reagent (GE Healthcare, Chicago, IL, USA) with the following antibodies: anti-FLAG (M2; Sigma-Aldrich), anti-pRb (4H1; Cell Signaling Technology, Danvers, MA), anti-PTPN14 (D5T6Y; Cell Signaling Technology), and anti-α-tubulin (B-5-1-2; Sigma-Aldrich). The band intensity of each protein was quantified with ChemiDoc XRS+ with Image Lab software (Bio-Rad, Hercules, CA, USA).
4
Western Blot Analysis of Protein Samples
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Monolayer cells were washed with 1× phosphate-buffered saline (PBS), harvested in 1.25× Laemmli buffer (78 mM Tris, pH 6.8; 2.5% SDS; 6.25% glycerol; 0.125% bromophenol blue; 2.5% β-mercaptoethanol), and immediately incubated at 99°C for 5 min and then placed on ice. Samples were treated with 100 U/ml Benzonase (Millipore) for 5 min at room temperature, and protein concentration was determined with a NanoDrop spectrophotometer (Thermo Fisher Scientific). Either 30 or 50 µg protein (as indicated in the figure legends) per lane was applied for Western blotting with the following antibodies: anti-FLAG M2 (Sigma-Aldrich catalog number F3165), anti-Strep-tag (IBA catalog number 2-1509-001), anti-p53 DO-1 (Santa Cruz Biotechnology catalog number sc-126), anti-pRb 4H1 (Cell Signaling Technology catalog number 9309), anti-HPV16 E7, clone NM2 (gift from Martin Müller, DKFZ), anti-HPV16 E6 (Euromedex catalog number E6-6F4), anti-kynureninase E-5 (Santa Cruz Biotechnology catalog number sc-390360), anti-CPPED1 H-11 (Santa Cruz Biotechnology catalog number sc-514222), anti-l- Plastin B-9 (Santa Cruz Biotechnology catalog number sc-133218), anti-transgelin 6G6 (Santa Cruz Biotechnology catalog number sc-53932), and goat-anti mouse IgG (H+L) horseradish peroxidase (HRP; Jackson ImmunoResearch).
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