Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

Acetyl-Histone H3 Antibody Sampler Kit #9927

acetyl histone   H3K14   H3K18   H3K27   H3K56   H3K9   Histone H3  

REACTIVITY

No. Size Price
9927T 1 Kit ( 6 x 20 µl ) ¥5,301.00 现货查询 购买询价
Kit Includes Quantity Applications Reactivity Homology† MW (kDa) Isotype
Acetyl-Histone H3 (Lys18) (D8Z5H) Rabbit mAb #13998 20 µl W,IP,IHC-P,F,ChIP, H,M,R,Mk,Sc, M,Hm,Pg, 17 Rabbit IgG
Acetyl-Histone H3 (Lys56) Antibody #4243 20 µl W, H,M,R,Mk, 17 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 (Lys14) (D4B9) Rabbit mAb #7627 20 µl W,IP,IF-IC,ChIP, H,M,R,Mk, Hm,Dm,X,Z,Pg,Sc,Hr, 17 Rabbit IgG
Acetyl-Histone H3 (Lys27) (D5E4) XP® Rabbit mAb #8173 20 µl W,IF-IC,F,ChIP, H,M,R,Mk, Hm,X,Z,GP,Hr, 17 Rabbit IgG
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

All antibodies in the Acetyl-Histone H3 Antibody Sampler Kit recognize histone H3 only when modified at the indicated site.

Acetyl-Histone H3 Antibody Sampler Kit的所有抗体仅识别指定位点修饰的histone H3蛋白。

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with synthetic acetylated peptides (KLH-coupled) corresponding to residues surrounding Lys9, Lys14, Lys18, Lys27, or Lys56 of human Histone H3. Antibodies are purified by protein A and peptide affinity chromatography.

通过合成的与人源Histone H3蛋白Lys9、Lys14、Lys18、Lys27或Lys56位点相应的乙酰化多肽(KLH-coupled)去免疫兔子从而制备出此多克隆抗体。通过蛋白A和多肽亲和层析纯化获得。

Description

The Acetyl-Histone H3 Antibody Sampler Kitprovides a fast and economical means of evaluating the acetylation sites on Histone H3. The kit contains enough primary and secondary antibodies to perform four Western mini-blot experiments.

Acetyl-Histone H3 Antibody Sampler Kit提供了一个快速经济的方法去评估Histone H3蛋白的乙酰化位点。该试剂盒包含充足的一抗和二抗去进行四次免疫印迹实验。

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 NIH/3T3 cells with or without TSA treatment, using Acetyl-Histone H3 (Lys18) Antibody #9675. 使用Acetyl-Histone H3 (Lys18) Antibody #9675,免疫印迹(Western blot)分析NIH/3T3细胞中Acetyl-Histone H3 (Lys18)蛋白水平,细胞分为untreated或TSA treated。

Western Blotting

Western Blotting

Western blot analysis of extracts from NIH/3T3 cells, untreated or TSA-treated, using Acetyl-Histone H3 (Lys18) Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human transitional epithelial carcinoma of the bladder showing nuclear localization using Acetyl-Histone H3 (Lys18) Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded NIH/3T3 cells, untreated (left) or TSA-treated (right), using Acetyl-Histone H3 (Lys18) Antibody. (no counterstain)

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.

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.

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 20 μl of Acetyl-Histone H3 (Lys18) Antibody 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.

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 HeLa, C6 and COS cells, untreated or treated with Trichostatin A (TSA) #9950 (400 nM for 18 h), using Acetyl-Histone H3 (Lys56) Antibody (upper) and Histone H3 Antibody #9715 (lower).

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left) and TSA-treated (#9950; right), using Acetyl-Histone H3 (Lys56) Antibody (green). Actin filaments were labeled with DY-554 phalloidin (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)分析不同细胞中Histone H3 (D1H2)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细胞中Acetyl-Histone H3 (Lys9)蛋白水平,细胞分为untreated或TSA-treated (400 nM for 18 hours)。

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa, C6 and COS cells, untreated or treated with Trichostatin A (TSA) #9950 (400 nM for 18 h), using Acetyl-Histone H3 (Lys56) Antibody #4243 (upper) and Histone H3 Antibody #9715 (lower). 使用Acetyl-Histone H3 (Lys56) Antibody #4243 (上图)和Histone H3 Antibody #9715 (下图),免疫印迹(Western blot)分析HeLa、C6和COS细胞中Acetyl-Histone H3 (Lys56)蛋白水平,细胞分为untreated或Trichostatin A (TSA) #9950 (400 nM for 18 h) treated。

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa and NIH/3T3 cells, untreated or treated with Trichostatin A #9950 (400 nM for 18 h), using Acetyl-Histone H3 (Lys14) Antibody #4318 (upper) and Histone H3 Antibody #9715 (lower). 使用Acetyl-Histone H3 (Lys14) Antibody #4318 (上图)和Histone H3 Antibody #9715 (下图),免疫印迹(Western blot)分析HeLa和NIH/3T3细胞中Acetyl-Histone H3 (Lys14)蛋白水平,细胞分为untreated或Trichostatin A (TSA) #9950 (400 nM for 18 h) treated。

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa and NIH/3T3 cells, untreated or treated with Trichostatin A #9950 (400 nM for 18 h), using Acetyl-Histone H3 (Lys27) Antibody #4353 (upper) and Histone H3 Antibody #9715 (lower). 使用Acetyl-Histone H3 (Lys27) Antibody #4353 (上图)和Histone H3 Antibody #9715 (下图),免疫印迹(Western blot)分析HeLa和NIH/3T3细胞中Acetyl-Histone H3 (Lys27)蛋白水平,细胞分为untreated或Trichostatin A (TSA) #9950 (400 nM for 18 h) treated。

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 (Lys14) (D4B9) 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 Intron 2 Primers #4478, 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.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa, C2C12, and COS-7 cells, untreated (-) or treated (+) with Trichostatin A (TSA) #9950 (1 μM, 18 h), using Acetyl-Histone H3 (Lys14) (D4B9) Rabbit mAb (upper) or Histone H3 (D1H2) XP® Rabbit mAb #4499 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa and C2C12 cells, untreated (-) or treated (+) with Trichostatin A (TSA) #9950 (1 μM, 18 hr), using Acetyl-Histone H3 (Lys27) (D5E4) XP® Rabbit mAb (upper) or Histone H3 (D1H2) XP® Rabbit mAb #4499 (lower).

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 5 μl of Acetyl-Histone H3 (Lys27) (D5E4) 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 AFM Intron 1 Primers #5098, 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 (left) or treated with Trichostatin A (TSA) #9950 (1 uM, 4 hr; right), using Acetyl-Histone H3 (Lys27) (D5E4) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).

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).

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.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded breast adenocarcinoma using Acetyl-Histone H3 (Lys18) (D8Z5H) Rabbit mAb in the presence of non-acetyl-Histone H3 peptide (left) or acetyl-Histone H3 (Lys18) peptide (right).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded transitional epithelial carcinoma of the bladder using Acetyl-Histone H3 (Lys18) (D8Z5H) 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, 18 hr), using Acetyl-Histone H3 (Lys18) (D8Z5H) Rabbit mAb (upper) and Histone H3 (D1H2) XP® Rabbit mAb #4499 (lower).

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 (Lys18) (D8Z5H) 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.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells using Acetyl-Histone H3 (Lys18) (D8Z5H) Rabbit mAb and Propidium Iodide (PI)/RNase Staining Solution #4087 to measure DNA content. Anti-rabbit IgG (H+L), F(ab)'2 Fragment (Alexa Fluor 488 Conjugate) #4412 was used as a secondary antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded colorectal adenocarcinoma using Acetyl-Histone H3 (Lys18) (D8Z5H) Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with Trichostatin A (TSA) #9950 (1 uM for 18 hours; right) using Acetyl-Histone H3 (Lys14) (D4B9) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HeLa cells, untreated (blue) or treated with Trichostatin A (TSA) #9950 (1 uM, Overnight; green) using Acetyl-Histone H3 (Lys27) (D5E4) XP® Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

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.

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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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