|H M R Mk||Endogenous||35-45||Rabbit|
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 datasheet or 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 November 2013
Reprobing of an existing membrane is a convenient means to immunoblot for multiple proteins independently when only a limited amount of sample is available. It should be noted that for the best possible results a fresh blot is always recommended. Reprobing can be a valuable method but with each reprobing of a blot there is potential for increased background signal. Additionally, it is recommended that you verify the removal of the first antibody complex prior to reprobing so that signal attributed to binding of the new antibody is not leftover signal from the first immunoblotting experiment. This can be done by re-exposing the blot to ECL reagents and making sure there is no signal prior to adding the next primary antibody.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalently purified water.
posted June 2005
revised June 2016
Protocol Id: 263
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.
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.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
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|8515S||100 µl (10 western blots)||$ 255.0|