Monoclonal Anti-TBP antibody produced in mouse

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	-Mouse (1:2000)

Protocol tips
-Mouse (1:2000)

Publication protocol

We lysed cells in RIPA buffer (10 mM Tris-HCL, 1 mM EDTA, 1% sodium dodecyl sulfate [SDS], 1% Nonidet P-40, 1: 100 proteinase inhibitor cocktail, 50 mM β-glycerophosphate, 50 mM sodium fluoride). We then separated lysates on a 10% SDS polyacrylamide gel and transferred to membranes by a semi-dry transfer apparatus (Bio-Rad). We blocked membranes with 5% milk for 1 h and then incubated with primary antibodies overnight. After rinsing, we incubated the immunocomplexes with horseradish peroxidase-conjugated anti-rabbit or anti-mouse IgG (Promega, Madison, WI) and visualized the membranes with SuperSignal Chemiluminiscent substrate (Pierce, Rockford, IL) as previously described22, 59. We purchased primary antibodies from the following commercial sources: anti-phospho-Tak1 (1:1000; 4531S; Cell Signaling, Danvers, Massachusetts), anti-Tak1 (1:1000; MAB5307; R&D systems), anti-phospho-p65 (1:2000; 3033S; Cell Signaling), anti-p65 (1:2000; 06-418; Millipore, Billerica, Massachusetts), anti-phospho-Iκbα (1:1000; 9246; Cell Signaling), anti-Iκbα (1:1000; sc-371; Santa Cruz, Santa Cruz, California), anti-phospho-JNK (1:500; 9251; Cell Signaling), anti-JNK (1:1,000; 9258; Cell Signaling), anti-phospho-p38 (1:1000; 9215; Cell Signaling), anti-p38 (1:1000; 8680; Cell Signaling), anti-phospho-Erk (1:1000; 4284; Cell Signaling), anti-Erk (1:1000; 4696; Cell Signaling), anti-Traf6 (2µg for immunoprecipitation; 1:1000 for Western blot; sc-8409; Santa Cruz), anti-Nlk (2µg for immunoprecipitation; AB10206, Millipore), anti-Tab2 (1:1000; 3744; Cell Signaling), anti-Nfatc1 (1:1000; sc-7294; Santa Cruz), anti-HA (1:2000; H9658; Sigma-Aldrich), anti-Tbp (1:2000; T1827; Sigma-Aldrich) and anti-α-tubulin (1:10000; 75168; Sigma-Aldrich).

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Western blotting TBP using Monoclonal Anti-TBP antibody produced in mouse from Sigma-Aldrich.

Manufacturer protocol

Download the product protocol from Sigma-Aldrich for Monoclonal Anti-TBP antibody produced in mouse below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Western blotting TBP using Monoclonal Anti-TBP antibody produced in mouse from Sigma-Aldrich. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.