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8686
Lysine Acetyltransferase Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Lysine Acetyltransferase Antibody Sampler Kit #8686

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Lysine Acetyltransferase Antibody Sampler Kit: Image 1
Western blot analysis of extracts from various cell lines using GCN5L2 (C26A10) Rabbit mAb.
Lysine Acetyltransferase Antibody Sampler Kit: Image 2
Western blot analysis of extracts from various cell lines using PCAF (C14G9) Rabbit mAb.
Lysine Acetyltransferase Antibody Sampler Kit: Image 3
Western blot analysis of extracts from A431, NIH/3T3, COS and PC12 cells, using Acetyl-CBP (Lys1535)/p300 (Lys1499) Antibody.
Lysine Acetyltransferase Antibody Sampler Kit: Image 4
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
Lysine Acetyltransferase Antibody Sampler Kit: Image 5
Western blot analysis of extracts from control HEK293T cells (Lane 1) or HEK293T cells with a targeted mutation in the gene encoding CBP (Lane 2) using CBP (D6C5) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). The change in CBP molecular weight in the mutated HEK293T cells confirms the specificity of the antibody for CBP.
Lysine Acetyltransferase Antibody Sampler Kit: Image 6
Confocal immunofluorescent analysis of HeLa cells using GCN5L2 (C26A10) Rabbit mAb (green). Actin filaments have been labeled with DY554 phalloidin (red). Blue pseudocolor = DRAQ5 (fluorescent DNA dye).
Lysine Acetyltransferase Antibody Sampler Kit: Image 7
Western blot analysis of hypo- or hyper-acetylated recombinant p300 HAT domains, either wild-type or K1499R mutant, using Acetyl-CBP (Lys1535)/p300 (Lys1499) Antibody (upper). Also shown in the corresponding coomassie stained SDS-PAGE gel (lower). (Details are described in Thompson, P.A. et al. (2004) Nat. Struct. Mol. Biol. 11, 308-315.)
Lysine Acetyltransferase Antibody Sampler Kit: Image 8
Western blot analysis of extracts from various cell lines using CBP (D6C5) Rabbit mAb.
Lysine Acetyltransferase Antibody Sampler Kit: Image 9
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® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. 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.
Lysine Acetyltransferase Antibody Sampler Kit: Image 10
Confocal immunofluorescent analysis of HeLa cells using CBP (D6C5) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Lysine Acetyltransferase Antibody Sampler Kit: Image 11
Chromatin immunoprecipitations were performed with cross-linked chromatin from 293 cells treated with Forskolin #3828 (30 μM, 1h) and CBP (D6C5) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across NR4A3, a known target gene of CBP (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.
Lysine Acetyltransferase Antibody Sampler Kit: Image 12
Chromatin immunoprecipitations were performed with cross-linked chromatin from 293 cells treated with Forskolin #3828 (30 μM, 1h) and CBP (D6C5) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across chromosome 9 (upper), including NR4A3 (lower), a known target gene of CBP (see additional figure containing ChIP-qPCR data).
Lysine Acetyltransferase Antibody Sampler Kit: Image 13
Chromatin immunoprecipitations were performed with cross-linked chromatin from 293 cells, treated with Forskolin #3828 (30 μM, 1h) and either CBP (D6C5) 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 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.
To Purchase # 8686
Cat. # Size Qty. Price
8686T
1 Kit  (4 x 20 microliters) $ 336

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
CBP (D6C5) Rabbit mAb 7389 20 µl
  • WB
  • IP
  • IF
  • ChIP
H M R Mk 300 Rabbit IgG
Acetyl-CBP (Lys1535)/p300 (Lys1499) Antibody 4771 20 µl
  • WB
  • IP
H M Mk 300 Rabbit 
PCAF (C14G9) Rabbit mAb 3378 20 µl
  • WB
  • IP
  • ChIP
H M R Mk 93 Rabbit IgG
GCN5L2 (C26A10) Rabbit mAb 3305 20 µl
  • WB
  • IP
  • IF
H M R Mk 94 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product 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 two western blots per primary.

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.

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.

Limited Uses

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