Revision 1

#12578

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877-616-CELL (2355)

orders@cellsignal.com

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, IP, ChIP, C&R

Reactivity:
H Mk

Sensitivity:
Endogenous

MW (kDa):
35-45

Source/Isotype:
Rabbit IgG

UniProt ID:
#P20226

Entrez-Gene Id:
6908

Product Usage Information

For optimal ChIP results, use 10 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 1:50
Chromatin IP 1:50
CUT&RUN 1:50

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 (D5G7Y) Rabbit mAb recognizes endogenous levels of total TBP protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala110 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.

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 nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

W: Western Blotting IP: Immunoprecipitation ChIP: Chromatin IP C&R: CUT&RUN

Cross-Reactivity Key

H: Human Mk: Monkey

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.

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Orders: 877-616-CELL (2355) orders@cellsignal.com Support: 877-678-TECH (8324) info@cellsignal.com Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using TBP (D5G7Y) Rabbit mAb.
Western Blotting Image 1: TBP (D5G7Y) Rabbit mAb
Immunoprecipitation of TBP from HCT 116 cell extracts, using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or TBP (D5G7Y) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using TBP (D5G7Y) Rabbit mAb.
Immunoprecipitation Image 1: TBP (D5G7Y) Rabbit mAb
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either TBP (D5G7Y) 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 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.
Chromatin Immunoprecipitation Image 1: TBP (D5G7Y) Rabbit mAb
Orders: 877-616-CELL (2355) orders@cellsignal.com Support: 877-678-TECH (8324) info@cellsignal.com Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
CUT&RUN was performed with HeLa cells and TBP (D5G7Y) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA library was prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across GAPDH, a known target gene of TBP (see additional figure containing CUT&RUN-qPCR data).
CUT & RUN Image 1: TBP (D5G7Y) Rabbit mAb
CUT&RUN was performed with HeLa cells and TBP (D5G7Y) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #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 & RUN Image 2: TBP (D5G7Y) Rabbit mAb
CUT&RUN was performed with HeLa cells and either TBP (D5G7Y) 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 γ-Actin Promoter Primers #5037, SimpleChIP® Human GAPDH Promoter Primers #4471 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 & RUN Image 3: TBP (D5G7Y) Rabbit mAb
Orders: 877-616-CELL (2355) orders@cellsignal.com Support: 877-678-TECH (8324) info@cellsignal.com Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.