Revision 7

#2316

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Orders:

877-616-CELL (2355)

[email protected]

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, IHC-P, IF-IC, FC-FP, C&R, eCLIP

Reactivity:
H M R Hm Mk B Dg

Sensitivity:
Endogenous

MW (kDa):
42, 55

Source/Isotype:
Rabbit IgG

UniProt ID:
#P50750

Entrez-Gene Id:
1025

Product Usage Information

The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.
Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Immunohistochemistry (Paraffin) 1:100 - 1:400
Immunofluorescence (Immunocytochemistry) 1:50 - 1:100
Flow Cytometry (Fixed/Permeabilized) 1:100 - 1:400
CUT&RUN 1:50
eCLIP 1:200
For more information about the RBP-eCLIP service please visit Eclipsebio.

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.

For a carrier free (BSA and azide free) version of this product see product #66968.

Specificity/Sensitivity

CDK9 (C12F7) Rabbit Monoclonal Antibody detects endogenous levels of total CDK9 protein, both 42 kDa and 55 kDa isoforms.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human CDK9.

Background

P-TEFb is a general transcription factor that regulates transcription elongation through phosphorylation of the C-terminal tail domain (CTD) of RNA polymerase II (RNAP II). The P-TEFb complex is composed of a catalytic subunit, CDK9, and its regulatory cyclin partner, which can be cyclin T1, T2a, T2b or K (reviewed in 1,2). P-TEFb is recruited by the HIV Tat protein to allow transcriptional elongation, and subsequent replication of the viral genome. Inhibition of P-TEFb function therefore has potential for HIV therapy. CDK9 exists as two isoforms, an abundant 42 kDa isoform, and a less abundant 55 kDa isoform, which contains an amino-terminal extension (3). The two forms likely have distinct purposes based on differential expression during lymphocyte activation (4,5) and on their localization within the nucleus (5).Cyclin dependent kinases (CDKs) are activated in part by cyclin binding and by phosphorylation of a conserved threonine in the T-loop domain. Phosphorylation of CDK9 at the T-loop Thr186 by an unidentified nuclear kinase may be important in P-TEFb activation (6) and regulation of HIV transcription (7). Acetylation of CDK9 at Lys44 affects its ability to phosphorylate the RNAPII CTD (8).

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

W: Western Blotting IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized) C&R: CUT&RUN eCLIP: eCLIP

Cross-Reactivity Key

H: Human M: Mouse R: Rat Hm: Hamster Mk: Monkey B: Bovine Dg: Dog

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Alexa Fluor is a registered trademark of Life Technologies Corporation.

U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 7

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell types using CDK9 (C12F7) Rabbit mAb.
Western Blotting Image 1: CDK9 (C12F7) Rabbit Monoclonal Antibody
Immunoprecipitation of CDK9 from HeLa cells using CDK9 (C12F7) Rabbit mAb. Western blot detection was performed using the same antibody. Lane 1 is 5% input.
Immunoprecipitation Image 1: CDK9 (C12F7) Rabbit Monoclonal Antibody
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using CDK9 (C12F7) Rabbit mAb in the presence of control peptide (left) or antigen specific peptide (right).
Immunohistochemistry Image 1: CDK9 (C12F7) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 7

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded K7M2 mouse syngeneic tumor using CDK9 (C12F7) Rabbit mAb.
Immunohistochemistry Image 2: CDK9 (C12F7) Rabbit Monoclonal Antibody
Confocal immunofluorescent analysis of HeLa cells using CDK9 (C12F7) Rabbit mAb (green). Actin filaments have been labeled with DY-555 phalloidin (red).
Immunofluorescence Image 1: CDK9 (C12F7) Rabbit Monoclonal Antibody
Flow cytometric analysis of HeLa cells using CDK9 (C12F7) Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Flow Cytometry Image 1: CDK9 (C12F7) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 7

Cell Signaling Technology Logo
CUT&RUN was performed with HeLa cells and CDK9 (C12F7) 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 the SLC25A25 gene.
CUT & RUN Image 1: CDK9 (C12F7) Rabbit Monoclonal Antibody
CUT&RUN was performed with HeLa cells and CDK9 (C12F7) 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 9 (upper), including the SLC25A25 gene (lower).
CUT & RUN Image 2: CDK9 (C12F7) Rabbit Monoclonal Antibody
CUT&RUN was performed with HeLa cells and either CDK9 (C12F7) 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 human SLC25A25 promoter primers and Myc intron 1 primers. 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: CDK9 (C12F7) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 7

Cell Signaling Technology Logo
Enhanced cross-linking and immunoprecipitation (eCLIP) was performed with RNA from Hep G2 cells and CDK9 (C12F7) Rabbit mAb using a protocol based on the RBP-eCLIP method from Eclipsebio. The figure shows binding across the PUM2 transcript. Data is kindly provided by the laboratory of Dr. Gene Yeo and used with permission.
eCLIP Image 1: CDK9 (C12F7) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.