Revision 4

#3378Store at -20C

Cell Signaling Technology

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Support: 877-678-TECH (8324)

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

WB, IP, ChIP, ChIP-seq

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

93

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q92831

Entrez-Gene Id:

8850

Product Information

Product Usage Information

For optimal ChIP and ChIP-seq results, use 20 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.

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Chromatin IP 1:25
Chromatin IP-seq 1:25

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

PCAF (C14G9) Rabbit mAb detects endogenous levels of total PCAF protein. The antibody does not cross-react with the related GCN5L2 protein.

Species Reactivity:

Human, Mouse, Rat, Monkey

Species predicted to react based on 100% sequence homology

Bovine, Horse

Source / Purification

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

Background

p300/CBP-associated factor (PCAF), also known as lysine acetyl-transferase 2B (KAT2B), is a transcriptional adaptor protein and histone acetyl-transferase (HAT) that functions as the catalytic subunit of the PCAF transcriptional co-activator complex (1). PCAF is 73% homologous to GCN5L2, another HAT protein found in similar complexes (1,2). Like GCN5L2, PCAF acetylates histone H3 on Lys14 and histone H4 on Lys8, both of which contribute to gene activation by modulating chromatin structure and recruiting additional co-activator proteins that contain acetyl-lysine binding bromo-domains (3). PCAF also acetylates non-histone proteins including transcriptional activators (p53, E2F1, MyoD), general transcription factors (TFIIEβ and TFIIF) and architectural DNA binding proteins (HMGA1 and HMG17) (4-10). Acetylation of these proteins regulates their nuclear localization, protein stability, DNA binding, and co-activator association.

  1. Nagy, Z. and Tora, L. (2007) Oncogene 26, 5341-57.
  2. Yang, X.J. et al. (1996) Nature 382, 319-24.
  3. Schiltz, R.L. et al. (1999) J Biol Chem 274, 1189-92.
  4. Bannister, A.J. and Miska, E.A. (2000) Cell Mol Life Sci 57, 1184-92.
  5. Liu, L. et al. (1999) Mol Cell Biol 19, 1202-9.
  6. Martínez-Balbás, M.A. et al. (2000) EMBO J 19, 662-71.
  7. Sartorelli, V. et al. (1999) Mol Cell 4, 725-34.
  8. Imhof, A. et al. (1997) Curr Biol 7, 689-92.
  9. Munshi, N. et al. (1998) Mol Cell 2, 457-67.
  10. Herrera, J.E. et al. (1999) Mol Cell Biol 19, 3466-73.

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

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.
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.

Revision 4
#3378

PCAF (C14G9) Rabbit mAb

Western Blotting Image 1: PCAF (C14G9) Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using PCAF (C14G9) Rabbit mAb.
No image available
Chromatin Immunoprecipitation Image 1: PCAF (C14G9) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from 293 cells treated with Forskolin #3828 (30 µM) for 1 hour and PCAF (C14G9) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figure shows binding across NR4A3, a known target gene of PCAF (see additional figure containing ChIP-qPCR data).
Chromatin Immunoprecipitation Image 2: PCAF (C14G9) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from 293 cells treated with Forskolin #3828 (30 µM) for 1 hour and PCAF (C14G9) Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 9 (upper), including NR4A3 (lower), a known target gene of PCAF (see additional figure containing ChIP-qPCR data).
Chromatin Immunoprecipitation Image 3: PCAF (C14G9) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from 293 cells treated with Forskolin #3828 (30 µM) and either PCAF (C14G9) Rabbit mAb or Normal Rabbit IgG #2729, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using human ALS2 exon 1 primers, SimpleChIP® Human NR4A3 Promoter Primers #4829, 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.