Revision 3

#2616Store at -20C

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, IHC-P, IF-IC, FC-FP, ChIP

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

25

SOURCE:

Rabbit

UniProt ID:

#P45973

Entrez-Gene Id:

23468

Product Information

Product Usage Information

For optimal ChIP 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 1:25
Immunohistochemistry (Paraffin) 1:200
Immunofluorescence (Immunocytochemistry) 1:200
Flow Cytometry (Fixed/Permeabilized) 1:100
Chromatin IP 1:25

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

HP1 alpha antibody detects endogenous levels of total HP1alpha protein. The antibody does not cross-react with HP1 beta or HP1 gamma proteins.

Species Reactivity:

Human, Mouse, Rat, Monkey

Species predicted to react based on 100% sequence homology

Bovine

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the carboxy terminus of human HP1alpha. Antibodies are purified by protein A and peptide affinity chromatography.

Background

Heterochromatin protein 1 (HP1) is a family of heterochromatic adaptor molecules involved in both gene silencing and higher order chromatin structure (1). All three HP1 family members (α, β, and γ) are primarily associated with centromeric heterochromatin; however, HP1β and γ also localize to euchromatic sites in the genome (2,3). HP1 proteins are approximately 25 kDa in size and contain a conserved amino-terminal chromodomain, followed by a variable hinge region and a conserved carboxy-terminal chromoshadow domain. The chromodomain facilitates binding to histone H3 tri-methylated at Lys9, a histone "mark" closely associated with centromeric heterochromatin (4,5). The variable hinge region binds both RNA and DNA in a sequence-independent manner (6). The chromoshadow domain mediates the dimerization of HP1 proteins, in addition to binding multiple proteins implicated in gene silencing and heterochromatin formation, including the SUV39H histone methyltransferase, the DNMT1 and DNMT3a DNA methyltransferases, and the p150 subunit of chromatin-assembly factor-1 (CAF1) (7-9). In addition to contributing to heterochromatin formation and propagation, HP1 and SUV39H are also found complexed with retinoblastoma (Rb) and E2F6 proteins, both of which function to repress euchromatic gene transcription in quiescent cells (10,11). HP1 proteins are subject to multiple types of post-translational modifications, including phosphorylation, acetylation, methylation, ubiquitination, and sumoylation, suggesting multiple means of regulation (12-14).

  1. Maison, C. and Almouzni, G. (2004) Nat. Rev. Mol. Cell Biol. 5, 296-304.
  2. Minc, E. et al. (2000) Cytogenet. Cell Genet. 90, 279-284.
  3. Nielsen, A.L. et al. (2001) Mol. Cell 7, 729-739.
  4. Lachner, M. et al. (2001) Nature 410, 116-120.
  5. Bannister, A.J. et al. (2001) Nature 410, 120-124.
  6. Muchardt, C. et al. (2002) EMBO Rep. 3, 975-981.
  7. Yamamoto, K. and Sonoda, M. (2003) Biochem. Biophys. Res. Commun. 301, 287-292.
  8. Fuks, F. et al. (2003) Nucleic Acids Res. 31, 2305-2312.
  9. Murzina, N. et al. (1999) Mol. Cell 4, 529-540.
  10. Nielsen, S.J. et al. (2001) Nature 412, 561-565.
  11. Ogawa, H. et al. (2002) Science 296, 1132-1136.
  12. Minc, E. et al. (1999) Chromosoma 108, 220-234.
  13. Zhao, T. et al. (2001) J. Biol. Chem. 276, 9512-9518.
  14. Lomberk, G. et al. (2006) Nat. Cell Biol. 8, 407-415.

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 IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized) ChIP: Chromatin IP

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.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

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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 3
#2616

HP1α Antibody

Western Blotting Image 1: HP1α Antibody Expand Image
使用 HP1alpha 抗体,对来自 HeLa 细胞、NIH/3T3 细胞、C6 细胞和 COS 细胞的提取物进行蛋白质印迹分析。
No image available
Immunohistochemistry Image 1: HP1α Antibody Expand Image
在对照肽(左)或 HP1 α 封闭肽 #1004(右)存在的情况下,使用 HP1alpha Antibody,对石蜡包埋的人乳腺癌进行免疫组织化学分析。
Immunofluorescence Image 1: HP1α Antibody Expand Image
使用 HP1α 抗体(绿色)标记的 HeLa 细胞进行共聚焦免疫荧光图像。肌动蛋白纤丝用 Alexa Fluor®555 phalloidin(红色)进行标记。
Flow Cytometry Image 1: HP1α Antibody Expand Image
使用与非特异性阴性对照抗体(红色)相对比的 HP1α 抗体(蓝色),对未处理的 HeLa 细胞进行流式细胞分析。
Chromatin Immunoprecipitation Image 1: HP1α Antibody Expand Image
使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003,对 mES 细胞中提取的交联染色质,在加入 HP1α Antibody #2616 或 Normal Rabbit IgG #2729 后,进行染色质免疫沉淀。使用 SimpleChIP® Mouse MEST Intron 1 Primers #12870、SimpleChIP® Mouse MEST Promoter Primers #12928、Mouse Intracisternal A-Particle (IAP) LTR 启动子引物和对非遗传性非重复区域特异的小鼠引物,通过实时 PCR 对富集的 DNA 进行定量分析。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。