Revision 4
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

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

WB, IP, FC-FP, ChIP, ChIP-seq

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

14

Source/Isotype:

Rabbit IgG

UniProt ID:

#P0C0S5

Entrez-Gene Id:

3015

Product Information

Product Usage Information

For optimal ChIP and ChIP-seq 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.

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:200
Flow Cytometry (Fixed/Permeabilized) 1:50
Chromatin IP 1:50
Chromatin IP-seq 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

Acetyl-Histone H2AZ (Lys4/Lys7) recognizes endogenous levels of histone H2AZ protein only when acetylated at Lys4 and/or Lys7. This antibody does not cross-react with other acetylated histones, including histone H2A acetylated at Lys5. This antibody also detects a band around 22 kDa, which is most likely monoubiquitylated histone H2AZ that is acetylated on Lys4 and Lys7.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding acetylated Lys4 and Lys7 of human H2AZ protein.

Background

Modulation of chromatin structure plays a critical role in the regulation of transcription in eukaryotes. The nucleosome, made up of four core histone proteins (H2A, H2B, H3 and H4), is the primary building block of chromatin. In addition to the growing number of post-translational histone modifications regulating chromatin structure, cells can also exchange canonical histones with variant histones that can directly or indirectly modulate chromatin structure (1). There are five major variants of histone H2A: canonical H2A (most abundant), H2A.X, MacroH2A, H2ABbd and H2A.Z (2). Histone H2A.Z, the most conserved variant across species, functions as both a positive and negative regulator of transcription and is important for chromosome stability (2). Several homologous protein complexes, such as SWR-C (S. cerevisiae), TIP60 (D. melanogaster) and SRCAP (mammals), have been shown to catalyze the ATP-dependent exchange of H2A.Z for H2A in the nucleosome (3,4,5). This exchange of histone H2A variants changes histone-histone interactions in the nucleosome core and alters an acidic patch on the surface of the nucleosome, resulting in changes in nucleosome stability and binding of non-histone proteins such as HP1α (6,7).

Acetylation of Histone H2AZ correlates with gene activity (8). Acetylation of Histone H2AZ on Lys4 and Lys7 occurs at the 5' end of genes and confers nucleome destabilization and open chromatin confirmation required for tanscriptional activation (9-11).

  1. Jin, J. et al. (2005) Trends Biochem Sci 30, 680-7.
  2. Raisner, R.M. and Madhani, H.D. (2006) Curr Opin Genet Dev 16, 119-24.
  3. Mizuguchi, G. et al. (2004) Science 303, 343-8.
  4. Kusch, T. et al. (2004) Science 306, 2084-7.
  5. Ruhl, D.D. et al. (2006) Biochemistry 45, 5671-7.
  6. Suto, R.K. et al. (2000) Nat Struct Biol 7, 1121-4.
  7. Fan, J.Y. et al. (2004) Mol Cell 16, 655-61.
  8. Millar, C.B. et al. (2006) Genes Dev 20, 711-22.
  9. Bruce, K. et al. (2005) Nucleic Acids Res 33, 5633-9.
  10. Ishibashi, T. et al. (2009) Biochemistry 48, 5007-17.
  11. Valdés-Mora, F. et al. (2012) Genome Res 22, 307-21.

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 FC-FP: Flow Cytometry (Fixed/Permeabilized) 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.
SimpleChIP 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.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Revision 4
#75336

Acetyl-Histone H2A.Z (Lys4/Lys7) (D3V1I) Rabbit mAb

Western Blotting Image 1: Acetyl-Histone H2A.Z (Lys4/Lys7) (D3V1I) Rabbit mAb Expand Image
使用 Acetyl-Histone H2A.Z (Lys4/Lys7) Rabbit mAb(上图)或 Histone H2A.Z Antibody #2718(下图)对未经处理 (-) 或经 Trichostatin A (TSA) #9950(1 μM,18 小时)处理 (+) 的 HeLa 细胞的提取物进行蛋白质印迹分析。
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Flow Cytometry Image 1: Acetyl-Histone H2A.Z (Lys4/Lys7) (D3V1I) Rabbit mAb Expand Image
使用 Acetyl-Histone H2A.Z (Lys4/Lys7) (D3V1I) Rabbit mAb 对未经处理(蓝色)或经 Trichostatin A (TSA) #9950(1 µM,18 小时,绿色)处理的 HeLa 细胞进行流式细胞分析。Anti-rabbit IgG (H+L)、F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 作为二抗。
Chromatin Immunoprecipitation Image 1: Acetyl-Histone H2A.Z (Lys4/Lys7) (D3V1I) Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005,以来自 Trichostatin A (TSA) #9950(100 nM,24 小时)处理的 LNCaP 细胞中的交联染色质和 Acetyl-Histone H2A.Z (Lys4/Lys7) (D3V1I) Rabbit mAb 进行染色质免疫沉淀。使用 DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795 制备 DNA 文库。该图图显示在 GAPDH 基因内结合。
Chromatin Immunoprecipitation Image 2: Acetyl-Histone H2A.Z (Lys4/Lys7) (D3V1I) Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005,以来自 Trichostatin A (TSA) #9950(100 nM,24 小时)处理的 LNCaP 细胞中的交联染色质和 Acetyl-Histone H2A.Z (Lys4/Lys7) (D3V1I) Rabbit mAb 进行染色质免疫沉淀。使用 DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795 制备 DNA 文库。该图显示在染色体 12(上图)内的结合,包括 GAPDH 基因(下图)。
Chromatin Immunoprecipitation Image 3: Acetyl-Histone H2A.Z (Lys4/Lys7) (D3V1I) Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005,以来自 Trichostatin A (TSA) #9950(100 nM,24 小时)处理的 LNCaP 细胞中的交联染色质和 Acetyl-Histone H2A.Z (Lys4/7) (D3V1I) Rabbit mAb 或 Normal Rabbit IgG #2729 进行染色质免疫沉淀。使用 Human CAV1 Promoter Primers、SimpleChIP® Human KLK2 Intron 1 Primers #62086和 SimpleChIP® Human α Satelite Repeat Primers #4486,通过实时 PCR 对富集的 DNA 进行定量。将每份样品中免疫沉淀的 DNA 的量表现为相对于所输入染色质总量(等于 1)的信号。