Antibodies used in immunoblotting: COL11A1 (ab64883, Abcam, 1:1000 dilution), ERα (No.8644, CST, 1:1000 dilution), anti-β-actin (60008-1-lg, Proteintech, 1:5000 dilution). Normal IgG/Peroxidase-conjugated AffiniPure Goat Anti-Rabbit/Mouse IgG (H + L) was purchased from Jackson Immuno Research. Tamoxifen metabolite 4-hydroxytamoxifen (4-OHT) was purchased from Sigma. Palbociclib HCl (#S1116), abemaciclib mesylate (#S7158), ribociclib HCl (#S5187) and fulvestrant (#S1191) were obtained from Selleck (Shanghai, China).
Anti β actin 60008 1 lg
Manufactured by Proteintech
Sourced in United States
Anti-β-actin (60008-1-lg) is a primary antibody that recognizes the beta-actin protein. Beta-actin is a highly conserved cytoskeletal protein found in all eukaryotic cells and is commonly used as a loading control in Western blotting experiments.
Automatically generated - may contain errors
Lab products found in correlation
Fulvestrant, by Selleck Chemicals (2 mentions)
Normal igg peroxidase conjugated affinipure goat anti rabbit mouse igg h l, by Jackson ImmunoResearch (2 mentions)
Ab64883, by Abcam (1 mentions)
Abemaciclib mesylate, by Selleck Chemicals (1 mentions)
Tamoxifen metabolite 4 hydroxytamoxifen 4 oht, by Merck Group (1 mentions)
Palbociclib hcl, by Selleck Chemicals (1 mentions)
Bca assay, by Beyotime (1 mentions)
Anti γh2ax ser139, by Merck Group (1 mentions)
Anti lig4 12695 1 ap, by Proteintech (1 mentions)
Imagequant las 500 system, by GE Healthcare (1 mentions)
Protease and phosphatase inhibitor, by Roche (1 mentions)
Anti p44 42 mapk erk1 2, by Cell Signaling Technology (1 mentions)
Anti akt, by Cell Signaling Technology (1 mentions)
Anti phospho erk1 2 thr202 tyr204, by Cell Signaling Technology (1 mentions)
Anti phospho akt, by Cell Signaling Technology (1 mentions)
Nitrocellulose membrane, by Bio-Rad (1 mentions)
Uchl1, by Cell Signaling Technology (1 mentions)
Cycloheximide (chx), by Avantor (1 mentions)
Anti gst 10000 0 ap, by Proteintech (1 mentions)
Ha no 3724, by Cell Signaling Technology (1 mentions)
4 protocols using anti β actin 60008 1 lg
1
Western Blot Antibody Validation
2
Western Blot Analysis of DNA Damage Response
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Cells were harvested in 0.25% trypsin (HyClone) and washed twice in ice-cold PBS. Total proteins were extracted in lysis buffer supplemented with protease and phosphatase inhibitors (Roche Applied Science). Protein concentrations were determined via BCA Assays (Beyotime Biotechnology). Samples were denatured in sodium dodecyl sulphate (SDS) sample buffer. Total protein extracts were separated by SDS-polyacrylamide gel electrophoresis and transferred to nitrocellulose membranes. Membranes were blocked in 5% nonfat dry milk in Tris-buffered saline-containing Tween-20 (TBST) and probed overnight at 4 °C with the indicated primary antibodies. The primary antibodies used in the study included: anti-β-Actin (60008-1-lg; Proteintech), anti-γ-H2AX ser139 (05-636; Millipore), and anti-LIG4 (12695-1-AP; Proteintech). Membranes were washed in TBST and labeled with the indicated horseradish peroxidase–conjugated secondary antibodies (KPL). Membranes were washed in TBST and protein bands were visualized using the Image Quant LAS500 system (GE Healthcare).
3
Western Blot Analysis of Signaling Proteins
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Proteins were separated on 7.5 or 10% sodium dodecyl sulfate-polyacrylamide gel and transferred to nitrocellulose membrane (Bio-Rad, Hercules, CA, United States). Then, the membrane was blocked with 5% non-fat milk and incubated with indicated primary antibodies. The proteins were detected using enhanced chemiluminescence reagents (Thermo Scientific). The primary antibodies used were anti-phospho-Akt (Ser473, #9271; Cell Signaling Technology, Beverly, MA, United States), anti-Akt (#9272, Cell Signaling Technology), anti-Phospho-Erk1/2 (Thr202/Tyr204, #4370, Cell Signaling Technology), anti-p44/42 MAPK (Erk1/2, #9102, Cell Signaling Technology), and anti-β-actin (60008-1-lg, Proteintech).
4
Dissecting EGFR Signaling Mechanisms
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Tamoxifen and MG132 were purchased from Sigma. Cycloheximide (CHX) was purchased from Amresco. LDN-57444 was purchased from Calbiochem. Fulvestrant and 17β-estradiol (E2) were purchased from Selleck. Antibodies used in immunoblotting: UCH-L1 (No.13179, 1:1000), ERα (No.8644, 1:1000), EGFR (No.4267, 1:1000), HA (No.3724, 1:1000) and pTyr1068-EGFR (No.3777, 1:1000) were purchased from Cell Signaling Technologies. Anti-GST (10000-0-AP, 1:4000), anti-Flag (66008-3-lg, 1:5000), anti-Myc (16286-1-AP, 1:2000), anti-UCH-L1 (66230-1-lg, 1:2000) and anti-β-actin (60008-1-lg, 1:5000) were purchased from Proteintech. Anti-eEF2K (ab45168, 1:1000) was purchased from Abcam. Anti-pThr678-EGFR (orb14895, 1:1000) was purchased from Biorbyt. Normal IgG/Peroxidase-conjugated AffiniPure Goat Anti-Rabbit/Mouse IgG (H+L) was purchased from Jackson Immuno Research.
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