Western blot analysis of extracts from various cell lines using TBP Antibody.
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Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 263
TBP Antibody recognizes endogenous levels of total TBP protein.
Human, Mouse, Rat, Monkey
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gln44 of human TBP protein. Antibodies are purified by protein A and peptide affinity chromatography.
TATA-binding protein (TBP) is a ubiquitously expressed nuclear protein that functions at the core of the general transcription factor protein complex TFIID (1-3). TFIID, which contains TBP and 13 TBP-associated factors (TAFs), contributes to the formation of the transcription pre-initiation complex, an assembly of multiple protein complexes (TFIIA, TFIIB, TFIIE, TFIIF, TFIIH, and RNA polymerase II) that bind to a gene promoter during the initiation of transcription (1-3). Once the pre-initiation complex is formed, RNA polymerase II becomes competent for elongation and transcribes the body of a gene. TBP functions in the recruitment of TFIID by binding to the TATA-box sequence found approximately 25 base pairs upstream of the transcription start site of many protein-coding genes. In addition, many transcriptional activator proteins interact with TBP and various TAF proteins to facilitate recruitment of TFIID and formation of the pre-initiation complex.
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