Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

Acetyl-Histone Antibody Sampler Kit #9933

h2a   h3   histone h3  

REACTIVITY

No. Size Price
9933T 1 Kit ( 8 x 20 µl ) ¥6,597.00 现货查询 购买询价
Kit Includes Quantity Applications Reactivity Homology† MW (kDa) Isotype
Histone H2A (D6O3A) Rabbit mAb #12349 20 µl W,IF-IC,ChIP, H,M,R,Mk,Z, Hm,B,Dg,GP, 14 Rabbit IgG
Histone H2B (D2H6) Rabbit mAb #12364 20 µl W,IHC-P,ChIP, H,M,R,Mk, C,Z,B, 14 Rabbit IgG
Acetyl-Histone H2B (Lys5) (D5H1S) XP® Rabbit mAb #12799 20 µl W,IP,IHC-P,IF-IC,ChIP, H,M,R,Mk, Hm,C,Z,B,Hr, 14 Rabbit IgG
Histone H4 (D2X4V) Rabbit mAb #13919 20 µl W,IHC-P,IF-IC, H,M,R,Mk, Hm,C,B, 11 Rabbit IgG
Acetyl-Histone H2A (Lys5) Antibody #2576 20 µl W,IP,IHC-P, H,M,R,Mk, 14 Rabbit
Acetyl-Histone H4 (Lys8) Antibody #2594 20 µl W, H,M,R,Mk, Ce, 11 Rabbit
Histone H3 (D1H2) XP® Rabbit mAb #4499 20 µl W,IHC-P,IF-IC,F, H,M,R,Mk, Hm,C,Dm,X,Z,B, 17 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl W, Goat
Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb #9649 20 µl W,IP,IHC-P,IF-IC,F,ChIP, H,M,R,Mk,Z,Ce, Sc, 17 Rabbit IgG

Specificity / Sensitivity

Each acetyl-histone antibody recognizes only the indicated protein target modified at the indicated site. Each control histone antibody recognizes the corresponding histone regardless of its acetylation state.

每一个acetyl-histone antibody仅可检测指定修饰位点的靶蛋白。无论它的乙酰化水平,每个对照组蛋白抗体都可识别相应的组蛋白。

Source / Purification

Polyclonal antibodies are produced by immunizing animals with synthetic acetylated peptides corresponding to residues surrounding Lys5 of histone H2A, Lys5 of histone H2B, Lys9 of histone H3, Lys8 of histone H4, or with synthetic peptides corresponding to the amino-terminal sequences of human histone H2A, H2B, or H4. Polyclonal Antibodies are purified by protein A and peptide affinity chromatography. Histone H3 monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the carboxy terminus of the human histone H3 protein. Acetyl-Histone H3 (Lys9) (C5B11) monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the amino terminus of histone H3 in which Lys9 is acetylated.

通过人工合成histone H2A蛋白Lys5位点、histone H2B蛋白Lys5位点、histone H3蛋白Lys9位点、histone H4蛋白Lys8位点周围相应的乙酰化多肽片段或人工合成人源histone H2A、H2B或H4蛋白氨基序列相应多肽片段去免疫动物从而制备出多克隆抗体。通过蛋白A和多肽亲和层析纯化抗体。Histone H3 monoclonal antibody是通过人工合成人源histone H3蛋白羧基端相应的多肽去免疫动物而制备。Acetyl-Histone H3 (Lys9) (C5B11) monoclonal antibody是通过人工合成histone H3蛋白氨基端Lys9位点乙酰化的多肽片段去免疫动物从而制备。

Description

The Acetyl-Histone Antibody Sampler Kit provides a fast and economical means of evaluating the acetylation states of histones H2A, H2B, H3 and H4. The kit contains enough primary and secondary antibodies to perform four Western blot experiments.

Acetyl-Histone Antibody Sampler Kit提供了一个快速经济的方法去测histones H2A、H2B、H3和H4蛋白的乙酰化水平。该试剂盒包含的一抗和二抗去做四次免疫印迹实验。

Western Blotting

Western Blotting

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.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines, untreated or TSA-treated (400 nM for 12 hours), using #2576 Acetyl-Histone H2A (Lys5) Antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines, untreated or TSA-treated (400 nM TSA for 12 hours), using Acetyl-Histone H4 (Lys8) Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical staining of paraffin-embedded human breast carcinoma, showing nuclear localization, using Acetyl-Histone H2A (Lys5) Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical staining of paraffin-embedded human breast carcinoma, showing nuclear localization of acetyl-histone H2A, using Acetyl-Histone H2A (Lys5) Antibody #2576.

使用Acetyl-Histone H2A (Lys5) Antibody #2576,免疫组化分析人源乳腺癌组织石蜡切片,显示acetyl-histone H2A的细胞核定位。

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines, untreated or TSA-treated (400 nM for 12 hours), using Acetyl-Histone H2A (Lys5) Antibody #2576.

使用Acetyl-Histone H2A (Lys5) Antibody #2576,免疫印迹(Western blot)分析不同细胞系中Acetyl-Histone H2A (Lys5)的蛋白水平,细胞分为未处理组或TSA(400 nM for 12 hours)处理组。

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines treated with TSA (400 nM for 12 hours) or untreated, using Acetyl-Histone H4 (Lys8) Antibody #2594.

使用Acetyl-Histone H4 (Lys8) Antibody #2594,免疫印迹(Western blot)分析不同细胞系中Acetyl-Histone H4 (Lys8)的蛋白水平,细胞分为未处理组或TSA(400 nM for 12 hours)处理组。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical staining of paraffin-embedded human breast carcinoma, showing nuclear localization of acetylated histone H4 (Lys8), using Acetyl-Histone H4 (Lys8) Antibody #2594.

使用Acetyl-Histone H4 (Lys8) Antibody #2594,免疫组化分析人源乳腺癌组织石蜡切片,结构显示acetylated histone H4 (Lys8)的细胞核定位。

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines, using Histone H4 Antibody #2592.

使用Histone H4 Antibody #2592,免疫印迹(Western blot)分析不同细胞系中Histone H4的蛋白水平。

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with TSA #9950 (right), using Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).

Western Blotting

Western Blotting

Western blot analysis of lysates from HeLa and NIH/3T3 cells, untreated or TSA-treated (400 nM for 18 hours) using Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb.

Western Blotting

Western Blotting

Western blot Analysis of extracts from NIH/3T3, HeLa and C6 cells using Histone H2A Antibody II #2578.

使用Histone H2A Antibody II #2578,免疫印迹(Western blot)分析NIH/3T3、HeLa和C6细胞系中Histone H2A的蛋白水平。

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines, untreated or TSA-treated (400 nM for 12 hours), using Acetyl-Histone H2B (Lys5) Antibody #2574.

使用Acetyl-Histone H2B (Lys5) Antibody #2574,免疫印迹(Western blot)分析不同细胞系中Acetyl-Histone H2B (Lys5)的蛋白水平,细胞分为未处理组或TSA(400 nM for 12 hours)处理组。

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 10 μl of Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human RPL30 Exon 3 Primers #7014, SimpleChIP® Human GAPDH Exon 1 Primers #5516, SimpleChIP® Human MyoD1 Exon 1 Primers #4490, 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.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Histone H4 (L64C1) Mouse mAb in the presence of control peptide (left) or antigen-specific peptide (right).

Western Blotting

Western Blotting

Western blot analysis of various cell lines using Histone H4 (L64C1) Mouse mAb.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of untreated HeLa cells using Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb #9649 versus propidium iodide (DNA content). Note positive staining in cycling cells (box).

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Histone H3 (D1H2) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Histone H3 (D1H2) XP® Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using Histone H3 (D1H2) XP® Rabbit mAb (green) and β-Tubulin (9F3) Rabbit mAb (Alexa Fluor® 555 Conjugate) #2116 (red).

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Histone H3 (D1H2) XP® Rabbit mAb #4499.

使用Histone H3 (D1H2) XP® Rabbit mAb #4499,免疫印迹(Western blot)分析不同细胞系中Histone H3 (D1H2)的蛋白水平。

Western Blotting

Western Blotting

Western blot analysis of lysates from HeLa and NIH/3T3 cells, untreated or TSA-treated (400 nM for 18 hours) using Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb #9649.

使用Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb #9649,免疫印迹(Western blot)分析HeLa和NIH/3T3细胞系中Acetyl-Histone H3 (Lys9)的蛋白水平,细胞分为未处理组或TSA(400 nM for 18 hours)处理组。

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Histone H2B (V119) Antibody #8135.

使用Histone H2B (V119) Antibody #8135,免疫印迹(Western blot)分析不同细胞系中Histone H2B (V119)的蛋白水平。

Western Blotting

Western Blotting

Western blot analysis of various cell lines using Histone H4 (L64C1) Mouse mAb #2935.

使用Histone H4 (L64C1) Mouse mAb #2935,免疫印迹(Western blot)分析不同细胞系中Histone H4 (L64C1)的蛋白水平。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human gastic carcinoma using Acetyl-Histone H3 (K9) Rabbit mAb in the presence of non-acetyl-peptide (left) or K9 acetyl-peptide (right).

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using Histone H2A (D6O3A) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Histone H2B (D2H6) Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Histone H2B (D2H6) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Histone H2A (D6O3A) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Histone H2B (D2H6) Rabbit mAb.

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 10 μl of Histone H2B (D2H6) Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. 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 MYT-1 Exon 1 Primers #4493, 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.

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 5 μl of Histone H2A (D6O3A) Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. 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 MYT-1 Exon 1 Primers #4493, 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.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (right) or treated with Trichostatin A (TSA) #9950 (left), using Acetyl-Histone H2B (Lys5) (D5H1S) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Acetyl-Histone H2B (Lys5) (D5H1S) XP® Rabbit mAb in the presence of non-acetyl peptide (left) or Lys5 acetyl peptide (right).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Acetyl-Histone H2B (Lys5) (D5H1S) XP® Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa and C6 cells, untreated (-) or treated with Trichostatin A (TSA) #9950 (1 μM, 18hr; +), using Acetyl-Histone H2B (Lys5) (D5H1S) XP® Rabbit mAb (upper) or Histone H2B (D2H6) Rabbit mAb #12364 (lower).

IP

IP

Immunoprecipitation of acetylated histone H2B from HeLa cell extracts treated with Trichostatin A (TSA) #9950 (1 μM, 18hr) using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Acetyl-Histone H2B (D5H1S) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Acetyl-Histone H2B (D5H1S) XP® Rabbit mAb.

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 10 µl of Acetyl-Histone H2B (Lys5) (D5H1S) XP® Rabbit mAb or 2 µl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. 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 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.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded mouse prostate using Histone H2B (D2H6) Rabbit mAb.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of human peripheral blood lymphocytes using Histone H3 (D1H2) XP® Rabbit mAb (blue) compared to Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414 was used as a secondary antibody.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using Histone H4 (D2X4V) Rabbit mAb (green) and Cox IV (4D11-B3-E8) Mouse mAb #11967 (red).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded mouse colon using Histone H4 (D2X4V) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using Histone H4 (D2X4V) Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Histone H4 (D2X4V) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast ductal carcinoma in situ using Histone H4 (D2X4V) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Histone H4 (D2X4V) Rabbit mAb.

Background

Modulation of chromatin structure plays an important role in the regulation of transcription in eukaryotes. The nucleosome, made up of DNA wound around eight core histone proteins (two each of H2A, H2B, H3, and H4), is the primary building block of chromatin (1). The amino-terminal tails of core histones undergo various post-translational modifications, including acetylation, phosphorylation, methylation, and ubiquitination (2-5). These modifications occur in response to various stimuli and have a direct effect on the accessibility of chromatin to transcription factors and, therefore, gene expression (6). In most species, histone H2B is primarily acetylated at Lys5, 12, 15, and 20 (4,7). Histone H3 is primarily acetylated at Lys9, 14, 18, 23, 27, and 56. Acetylation of H3 at Lys9 appears to have a dominant role in histone deposition and chromatin assembly in some organisms (2,3). Phosphorylation at Ser10, Ser28, and Thr11 of histone H3 is tightly correlated with chromosome condensation during both mitosis and meiosis (8-10). Phosphorylation at Thr3 of histone H3 is highly conserved among many species and is catalyzed by the kinase haspin. Immunostaining with phospho-specific antibodies in mammalian cells reveals mitotic phosphorylation at Thr3 of H3 in prophase and its dephosphorylation during anaphase (11).

染色质结构的修饰在调节真核细胞的转录中扮演着重要的角色。由DNA围绕和八聚体组蛋白(H2A,H2B,H3和H4各两个)共同组成的核小体是染色质的主要组成(1)。核小体的组蛋白氨基酸尾端进行不同的转录后修饰,包括乙酰化,磷酸化,甲基化和泛素化(2-5)。这些修饰通过不同的刺激产生并且对转录因子能否接近染色质有着直接的影响,所以也影响着基因的表达(6)。在大多数的物种中组蛋白H2B主要在Lys5,,12,,15和20位点上发生乙酰化(4,7)。组蛋白H3主要是在Lys9,14,18,23,27和56位点上发生乙酰化。在某些物种里H3上的Lys9位点发生乙酰化并应该在组蛋白沉积和染色质组装中扮演着重要的角色(2,3)。组蛋白H3上Ser10,Ser28和Thr11位点的磷酸化在有丝分裂和无丝分裂中都与染色质的缩合紧密相连(8-10)。H3的Thr3位点的磷酸化在许多物种中都是高度保守的,是由kinase haspin所催化的。在哺乳动物中用磷酸化特异性的抗体做免疫组化显示H3的Thr3位点在有丝分裂的前期发生磷酸化,后期发生去磷酸化(11)。

  1. Workman, J.L. and Kingston, R.E. (1998) Annu Rev Biochem 67, 545-79.
  2. Hansen, J.C. et al. (1998) Biochemistry 37, 17637-41.
  3. Strahl, B.D. and Allis, C.D. (2000) Nature 403, 41-5.
  4. Cheung, P. et al. (2000) Cell 103, 263-71.
  5. Bernstein, B.E. and Schreiber, S.L. (2002) Chem Biol 9, 1167-73.
  6. Jaskelioff, M. and Peterson, C.L. (2003) Nat Cell Biol 5, 395-9.
  7. Thorne, A.W. et al. (1990) Eur J Biochem 193, 701-13.
  8. Hendzel, M.J. et al. (1997) Chromosoma 106, 348-60.
  9. Goto, H. et al. (1999) J Biol Chem 274, 25543-9.
  10. Preuss, U. et al. (2003) Nucleic Acids Res 31, 878-85.
  11. Dai, J. et al. (2005) Genes Dev 19, 472-88.

Application References

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

U.S. Patent No. 5,675,063.

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

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

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