Cell Signaling Technology

Product Pathways - Motif Antibodies

Acetylated-Lysine Antibody #9441

acetyl lysine   acetyl-histone   acetylated lysine   acetylation   general   histone   motif   sc-377521   substrate  

No. Size Price
9441L 300 µl ( 30 western blots ) ¥8,792.00 现货查询 购买询价
9441S 100 µl ( 10 western blots ) ¥3,900.00 现货查询 购买询价
9441 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey,S. cerevisiae,All Species Expected, Endogenous Rabbit
IP 1:100
IHC-P 1:800
IF-IC 1:100
ChIP 1:50
E-P 1:2000

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), IF-IC=Immunofluorescence (Immunocytochemistry), ChIP=Chromatin IP, E-P=Peptide ELISA (DELFIA),

Specificity / Sensitivity

Acetylated-Lysine Antibody detects proteins posttranslationally modified by acetylation on the epsilon-amine groups of lysine residues. The antibody recognizes acetylated lysine in a wide range of sequence contexts. It has been demonstrated to recognize acetylated histones, p53, CBP, PCAF and chemically acetylated BSA. The antibody has been shown to react with as little as 0.04 ng of chemically acetylated BSA while not recognizing up to 25 µg of nonacetylated BSA. (U.S. Patent No's.: 6,441,140; 6,982,318; 7,259,022; 7,344,714; U.S.S.N. 11,484,485; and all foreign equivalents.)

Acetylated-Lysine Antibody乙酰化赖氨酸的兔多抗识别转录后修饰为赖氨酸残基ε-氨基乙酰化的蛋白质。此抗体可识别不同背景序列中的乙酰化赖氨酸。研究显示它可以识别乙酰化的组蛋白、p53、CBP、PCAF和化学乙酰化的BSA。研究显示抗体可以与少到0.04ng的化学乙酰化的BSA反应,却不能与非乙酰化的多到25ug的BSA反应。(美国专利号:6,441,140、6,982,318、7,259,022、7,344,714;U.S.S.N. 11,484,485;及所有国外相应专利)

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic acetylated lysine-containing peptide. Antibodies are purified by protein A and peptide affinity chromatography.

该多克隆抗体用合成的含乙酰化赖氨酸的肽段免疫动物制备。该抗体使用蛋白A和蛋白亲和层析纯化而得。

Western Blotting

Western Blotting

Western blot analysis of extracts from NIH/3T3 cells, untreated or sodium butyrate-treated (5 mM for 24 hours), showing an increase in histone acetylation using Acetylated-Lysine Antibody.

对NIH/3T3细胞抽提液,未处理或5 mM丁酸钠处理24小时,使用Acetylated-Lysine Antibody进行Western blot分析,显示组蛋白乙酰化增加。

IP

IP

Western blot analysis of immunoprecipitated p53 showing an increase in p53 acetylation using Acetylated-Lysine Antibody (upper) or p53 antibody (lower). p53 was immunoprecipitated from lysates from 293 cells, untreated or UV-treated, using p53 Antibody #9282.

对免疫共沉淀的p53,使用Acetylated-Lysine Antibody(上图)或p53 antibody(下图)进行Western blot分析,显示p53乙酰化增加。从293细胞裂解后免疫共沉淀的p53,未处理或UV处理,使用p53 Antibody #9282。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical staining of a paraffin-embedded human breast tumor section showing nuclear and cytoplasmic localization of proteins with acetylated lysine residues using Acetylated-Lysine Antibody.

对石蜡包埋的人乳腺肿瘤切片使用Acetylated-Lysine Antibody进行免疫组化分析,显示细胞核和细胞质定位。

Western Blotting

Western Blotting

Western blot analysis of extracts from COS cells, untreated or TSA-treated, grown in 10% FBS (lanes 1 and 2) or serum starved for 18 hours (lanes 3 and 4), using Acetylated-Lysine Antibody (upper) or p44/42 MAP Kinase Antibody #9102 (lower).

对COS细胞,未处理或TSA处理,在10% FBS(列1和列2)生长或血清饥饿处理18小时(列3和列4),使用Acetylated-Lysine Antibody(上图)或p44/42 MAP Kinase Antibody #9102(下图)进行Western blot分析。

Western Blotting

Western Blotting

Specificity and sensitivity of Acetylated-Lysine Antibody assayed on acetylated BSA (4; 1; 0.2; 0.04 or 0.008 ng in lanes 1-5) or nonacetylated BSA (25,000; 5,000; 1,000 or 200 ng in lanes 6-9).

Acetylated-Lysine Antibody的特异性或敏感性用乙酰化BSA(4、1、0.2、0.04或0.008 ng 列1-5)和非乙酰化BSA(25,000、5,000、1,000或200 ng 列6-9)进行检测。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded NIH/3T3 untreated (left) or TSA-treated (right) using Acetylated-Lysine Antibody.

对NIH/3T3细胞,未处理(左)或TSA处理(右),使用Acetylated-Lysine Antibody进行免疫组化分析。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Acetylated-Lysine Antibody.

对石蜡包埋的人结肠癌使用Acetylated-Lysine Antibody抗体进行免疫组化分析。

IF-IC

IF-IC

Confocal immunofluorescent analysis of NIH/3T3 cells, untreated (left) or SAHA-treated (right), labeled with Acetylated-Lysine Antibody (green). Actin filaments have been labeled with Alexa Fluor R 555 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

对NIH/3T3细胞,未处理(左)或SAHA处理(右),使用Acetylated-Lysine Antibody(绿)进行共聚焦免疫荧光分析。肌动蛋白用Alexa Fluor® 555 phalloidin(红色)标记。蓝色伪彩=DRAQ5®#4084(荧光DNA染料)。

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 20 μl of Acetylated-Lysine Antibody or 2 μl of Normal Rabbit IgG #2729, using SimpleChIP® Enzymatic Chromatin IP Kit (Agarose Beads) #9002. The enriched DNA was quantified by real-time PCR, using SimpleChIP® Human GAPDH Exon 1 Primers #5516, SimpleChIP® Human RPL30 Exon 3 Primers #7014, SimpleChIP® Human MyoD1 Exon 1 Primers #4490, and SimpleChIP® Human MYT-1 Exon 1 Primers #4493. 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.

使用SimpleChIP® Enzymatic Chromatin IP Kit (Agarose Beads) #9002试剂盒,将4 x 10^6 HeLa细胞的染色质,与20μl Acetylated-Lysine (Ac-K2-100) Rabbit mAb #9814或者2 μl of Normal Rabbit IgG #2729进行染色质免疫沉淀。使用人SimpleChIP® Human GAPDH Exon 1 Primers #5516, SimpleChIP® Human RPL30 Exon 3 Primers #7014, SimpleChIP® Human MyoD1 Exon 1 Primers #4490和SimpleChIP® Human MYT-1 Exon 1 Primers #4493进行real-time PCR用来定量富集的DNA。每个样本中免疫沉淀的DNA使用相对于输入对照中染色质的量描述,输入对照的量相当于1。

Background

Acetylation of lysine, like phosphorylation of serine, threonine or tyrosine, is an important reversible modification controlling protein activity. The conserved amino-terminal domains of the four core histones (H2A, H2B, H3, and H4) contain lysines that are acetylated by histone acetyltransferases (HATs) and deacetylated by histone deacetylases (HDACs) (1). Signaling resulting in acetylation/deacetylation of histones, transcription factors, and other proteins affects a diverse array of cellular processes including chromatin structure and gene activity, cell growth, differentiation, and apoptosis (2-6). Recent proteomic surveys suggest that acetylation of lysine residues may be a widespread and important form of posttranslational protein modification that affects thousands of proteins involved in control of cell cycle and metabolism, longevity, actin polymerization, and nuclear transport (7,8). The regulation of protein acetylation status is impaired in cancer and polyglutamine diseases (9), and HDACs have become promising targets for anti-cancer drugs currently in development (10).

赖氨酸的乙酰化如同丝氨酸、苏氨酸和酪氨酸的磷酸化一样,是一种控制蛋白活性的重要可逆手段。四个核心组蛋白(H2A, H2B, H3, H4)的保守氨基末端区域均包含赖氨酸,可以被组蛋白乙酰转移酶(HATs)乙酰化和组蛋白去乙酰酶(HDACs)去乙酰化(1)。组蛋白、转录因子和其它蛋白的乙酰化/去乙酰化信号最终影响多个细胞过程,包括染色体结构、基因活性、细胞生长、分化和凋亡等(2-6)。最近的蛋白组学研究显示赖氨酸残基的乙酰化是蛋白翻译后修饰的广泛而重要的手段,可以影响控制细胞周期、代谢、生存、肌动蛋白聚合和核转运相关的数千种蛋白。蛋白质乙酰化调节障碍在癌症和多聚谷氨酰胺疾病中发现(9),HDACs也成为目前正在开发的有希望的抗癌药物靶点(10)。

  1. Hassig, C.A. and Schreiber, S.L. (1997) Curr Opin Chem Biol 1, 300-8.
  2. Allfrey, V.G. et al. (1964) Proc Natl Acad Sci USA 51, 786-94.
  3. Liu, L. et al. (1999) Mol Cell Biol 19, 1202-9.
  4. Boyes, J. et al. (1998) Nature 396, 594-8.
  5. Polevoda, B. and Sherman, F. (2002) Genome Biol 3, reviews 0006.
  6. Yoshida, M. et al. (2003) Prog Cell Cycle Res 5, 269-78.
  7. Kim, S.C. et al. (2006) Mol Cell 23, 607-18.
  8. Choudhary, C. et al. (2009) Science 325, 834-40.
  9. Hughes, R.E. (2002) Curr Biol 12, R141-3.
  10. Vigushin, D.M. and Coombes, R.C. (2004) Curr Cancer Drug Targets 4, 205-18.

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

Use of Cell Signaling Technology (CST) Motif Antibodies within certain methods (e.g., U.S. Patents No. 7,198,896 and 7,300,753) may require a license from CST. For information regarding academic licensing terms please have your technology transfer office contact CST Legal Department at CST_ip@cellsignal.com. For information regarding commercial licensing terms please contact CST Pharma Services Department at ptmscan@cellsignal.com.

DRAQ5 is a registered trademark of Biostatus Limited.

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

SignalStain is a trademark of Cell Signaling Technology, Inc.

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

Alexa Fluor is a registered trademark of Life Technologies Corporation.

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

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