Product Pathways - Chromatin Regulation / Epigenetics

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Lysine Acetyltransferase Antibody Sampler Kit #8686

Kit Includes	Quantity	Applications	Reactivity	MW (kDa)	Isotype
CBP (D6C5) Rabbit mAb #7389	40 µl	W IP IF-IC ChIP	H M R Mk	300	Rabbit IgG
Acetyl-CBP (Lys1535)/p300 (Lys1499) Antibody #4771	40 µl	W IP ChIP	H M Mk (R)	300	Rabbit
PCAF (C14G9) Rabbit mAb #3378	40 µl	W IP ChIP	H M R Mk (B) (Hr)	93	Rabbit IgG
GCN5L2 (C26A10) Rabbit mAb #3305	40 µl	W IP IF-IC	H M R Mk (B) (Hr)	94	Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074	100 µl				Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)  ChIP=Chromatin IP
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  B=Bovine  Hr=Horse
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

Each antibody in the Lysine Acetyltransferase Antibody Sampler Kit recognizes endogenous levels of respective target protein. The antibodies do not cross-react with other family members.

Description

The Lysine Acetyltransferase Antibody Sampler Kit provides an economical means to examine several lysine acetyltrasferases, including: Acetyl-CBP, CBP, GCN5L2, and PCAF. The kit contains enough primary antibody to perform four western blots per primary.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the amino terminus of human CBP protein, human GCN5L2 protein, or human PCAF protein. Polyclonal antibodies are produced by immunizing animals with a synthetic acetylated peptide corresponding to residues surrounding Lys1535 of human CBP. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

Background

CREB-binding protein (CBP) and p300 are highly conserved and functionally related transcriptional co-activators that associate with transcriptional regulators and signaling molecules, integrating multiple signal transduction pathways with the transcriptional machinery (1,2). CBP/p300 also contain histone acetyltransferase (HAT) activity, allowing them to acetylate histones and other proteins (2). The role of acetylation of CBP/p300 is of particular interest (2,3). Acetylation of p300 at Lys1499 has been demonstrated to enhance its HAT activity and affect a wide variety of signaling events (4). p300/CBP-associated factor (PCAF), also known as lysine acetyl-transferase 2B (KAT2B) (5), and General Control of Amino Acid Synthesis Yeast Homolog Like 2 (GCN5L2) (6) are transcriptional adaptor proteins in addition to HATs. PCAF functions as the catalytic subunit of the PCAF transcriptional co-activator complex (5). GCN5L2 functions as the catalytic subunit of the STAGA and TFTC transcription coactivator complexes (6). PCAF and GCN5L2 acetylate histone H3 at Lys14 and histone H4 at 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 (7,8). PCAF also acetylates non-histone proteins including transcriptional activators (p53, E2F1, MyoD) and general transcription factors (TFIIEβ and TFIIF) (9-12). GCN5L2 also acetylates non-histone proteins such as transcription activators (TAT, c-Myb) (13,14), transcription co-activators (PGC1-α) (15), and nuclear receptors (Steroidogenic Factor 1) (16). Acetylation of these proteins regulates their nuclear localization, protein stability, DNA binding, and co-activator association (13-16).

1. Goodman, R.H. and Smolik, S. (2000) Genes Dev 14, 1553-77.
2. Chan, H.M. and La Thangue, N.B. (2001) J Cell Sci 114, 2363-73.
3. Yuan, L.W. and Giordano, A. (2002) Oncogene 21, 2253-60.
4. Thompson, P.R. et al. (2004) Nat Struct Mol Biol 11, 308-15.
5. Nagy, Z. and Tora, L. (2007) Oncogene 26, 5341-57.
6. Candau, R. et al. (1996) Mol Cell Biol 16, 593-602.
7. Schiltz, R.L. et al. (1999) J Biol Chem 274, 1189-92.
8. Grant, P.A. et al. (1999) J Biol Chem 274, 5895-900.
9. Bannister, A.J. and Miska, E.A. (2000) Cell Mol Life Sci 57, 1184-92.
10. Liu, L. et al. (1999) Mol Cell Biol 19, 1202-9.
11. Sartorelli, V. et al. (1999) Mol Cell 4, 725-34.
12. Imhof, A. et al. (1997) Curr Biol 7, 689-92.
13. Kiernan, R.E. et al. (1999) EMBO J 18, 6106-18.
14. Tomita, A. et al. (2000) Oncogene 19, 444-51.
15. Lerin, C. et al. (2006) Cell Metab 3, 429-38.
16. Jacob, A.L. et al. (2001) J Biol Chem 276, 37659-64.

Application References

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Protocols

3305:

Immunofluorescence, Immunoprecipitation, Western Blotting

3378:

ChIP Agarose, ChIP Magnetic, Immunoprecipitation, Western Blotting

4771:

ChIP Agarose, ChIP Magnetic, Immunoprecipitation, Western Blotting

7389:

Immunofluorescence, Immunoprecipitation, Western Blotting

Selected rabbit monoclonal antibodies are produced under license (granting certain rights including those under U. S. Patents No. 5,675,063) from Epitomics, Inc. 

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For Research Use Only. Not For Use In Diagnostic Procedures.