Revision 5

#44059Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, ChIP, ChIP-seq, C&R

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

35-45

Source/Isotype:

Rabbit IgG

UniProt ID:

#P20226

Entrez-Gene Id:

6908

Product Information

Product Usage Information

For optimal ChIP and ChIP-seq results, use 5 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.
Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Chromatin IP 1:100
Chromatin IP-seq 1:100
CUT&RUN 1:100

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

TBP (D5C9H) XP® Rabbit mAb recognizes endogenous levels of total TBP protein.

Species Reactivity:

Human, Mouse, Rat, Monkey

Species predicted to react based on 100% sequence homology

Chicken, Bovine

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human TBP protein.

Background

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.

  1. Goodrich, J.A. and Tjian, R. (1994) Curr Opin Cell Biol 6, 403-9.
  2. Berk, A.J. (2000) Cell 103, 5-8.
  3. Thomas, M.C. and Chiang, C.M. (2006) Crit Rev Biochem Mol Biol 41, 105-78.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq C&R: CUT&RUN

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

使用に関する制限

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Revision 5
#44059

TBP (D5C9H) XP® Rabbit mAb

Western Blotting Image 1: TBP (D5C9H) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using TBP (D5C9H) XP® Rabbit mAb.
Immunoprecipitation Image 1: TBP (D5C9H) XP® Rabbit mAb Expand Image
Immunoprecipitation of TBP from HeLa cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is TBP (D5C9H) XP® Rabbit mAb. Western blot analysis was performed using TBP (D5C9H) XP® Rabbit mAb.
Chromatin Immunoprecipitation Image 1: TBP (D5C9H) XP® Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and TBP (D5C9H) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across GAPDH, a known target gene of TBP (see additional figure containing ChIP-qPCR data).
Chromatin Immunoprecipitation Image 2: TBP (D5C9H) XP® Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and TBP (D5C9H) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 12 (upper), including GAPDH (lower), a known target gene of TBP (see additional figure containing ChIP-qPCR data).
Chromatin Immunoprecipitation Image 3: TBP (D5C9H) XP® Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either TBP (D5C9H) XP® Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human γ-Actin Promoter Primers #5037, SimpleChIP® Human GAPDH Promoter Primers #4471, SimpleChIP® Human MYT-1 Exon 1 Primers #4493, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
CUT and RUN Image 1: TBP (D5C9H) XP® Rabbit mAb Expand Image
CUT&RUN was performed with HeLa cells and TBP (D5C9H) XP® Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Library was prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across GAPDH, a known target gene of TBP (see additional figure containing CUT&RUN-qPCR data).
CUT and RUN Image 2: TBP (D5C9H) XP® Rabbit mAb Expand Image
CUT&RUN was performed with HeLa cells and TBP (D5C9H) XP® Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Library was prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 12 (upper), including GAPDH (lower), a known target gene of TBP (see additional figure containing CUT&RUN-qPCR data).
CUT and RUN Image 3: TBP (D5C9H) XP® Rabbit mAb Expand Image
CUT&RUN was performed with HeLa cells and either TBP (D5C9H) XP® Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human GAPDH Promoter Primers #4471, SimpleChIP® Human γ-Actin Promoter Primers #5037, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.