Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

Acetyl-Histone H4 Antibody Sampler Kit #8346

REACTIVITY

No. Size Price
8346T 1 Kit ( 4 x 20 µl ) ¥3,876.00 现货查询 购买询价
Kit Includes Quantity Applications Reactivity Homology† MW (kDa) Isotype
Acetyl-Histone H4 (Lys12) (D2W6O) Rabbit mAb #13944 20 µl W,IP,IF-IC,F,ChIP, H,R,Mk, C,Dm, 11 Rabbit IgG
Acetyl-Histone H4 (Lys8) Antibody #2594 20 µl W, H,M,R,Mk, Ce, 11 Rabbit
Histone H4 (L64C1) Mouse mAb #2935 20 µl W,IHC-P, H,M,R,Mk, Dm,X,Z,B,Hr,Ce, 11 Mouse IgG1
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl W, Goat
Anti-mouse IgG, HRP-linked Antibody #7076 100 µl W, Horse
Acetyl-Histone H4 (Lys5) (D12B3) Rabbit mAb #8647 20 µl W,IP,IHC-P,IF-IC,ChIP, H,M,R,Mk, C,Dm,X,Z,B,Pg,Hr,Ce, 11 Rabbit IgG

Specificity / Sensitivity

Each antibody in this kit recognizes only its specific target protein and does not cross-react with other family members.

每一个抗体仅可检测特异性靶蛋白,并且不能与其它家族成员发生交叉反应。

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic acetylated peptide corresponding to residues surrounding Lys12, Lys5 or Lys8 of human histone H4. Antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the amino-terminal sequence of human histone H4.

通过人工合成人源histone H4蛋白Lys12、Lys5或Lys8位点周围相应的乙酰化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获得。通过人工合成人源histone H4蛋白氨基端序列去免疫动物从而制备出单克隆抗体。

Description

The Acetyl-Histone H4 Antibody Sampler Kit provides an economical means of detecting total histone H4 as well as histone H4 acetylated at various residues including Lys12, Lys5, and Lys8. The kit contains enough primary and secondary antibody to perform four western blots with each antibody.

Acetyl-Histone H4 Antibody Sampler Kit提供了一个经济的方法去检测histone H4总蛋白和在Lys12、Lys5和Lys8位点乙酰化的histone H4蛋白。该试剂盒包含的一抗和二抗可以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

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 TSA for 12 hours) using Acetyl-Histone H4 (Lys12) 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 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.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or TSA-treated (right), using Acetyl-Histone H4 (Lys12) Antibody (green). Actin filaments have been labled with DY-554 phalloidin (red).

Western blot analysis of extracts from various cell lines, untreated or TSA-treated (400 nM TSA for 12 hours), using Acetyl-Histone H4 (Lys12) Antibody #2591.

使用Acetyl-Histone H4 (Lys12) Antibody #2591,免疫印迹(Western blot)分析不同细胞系中Acetyl-Histone H4 (Lys12)的蛋白水平,细胞分为未处理组或TSA(400 nM for 12 hours)处理组。

Western blot analysis of extracts from various cell lines using Acetyl-Histone H4 (Lys5) Antibody #9672.

使用Acetyl-Histone H4 (Lys5) Antibody #9672,免疫印迹(Western blot)分析不同细胞系中Acetyl-Histone H4 (Lys5)的蛋白水平。

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 #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。

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。

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with Trichostatin A (TSA) #9950 (right) using Acetyl-Histone H4 (Lys5) (D12B3) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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 H4 (Lys5) (D12B3) 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.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Acetyl-Histone H4 (Lys5) (D12B3) Rabbit mAb.

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 H4 (Lys5) (D12B3) Rabbit mAb (upper) or Histone H4 (L64C1) Mouse mAb #2935 (lower).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast using Acetyl-Histone H4 (Lys5) (D12B3) Rabbit mAb in the presence of non-acetyl-histone H4 (Lys5) peptide (left) or acetyl-histone H4 (Lys5) peptide (right).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Acetyl-Histone H4 (Lys5) (D12B3) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Acetyl-Histone H4 (Lys5) (D12B3) 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 H4 (Lys12) (D2W6O) Rabbit mAb (upper) and Histone H4 (D2X4V) Rabbit mAb #13919 (lower).

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells untreated (left) or treated (right) with Trichostatin A (TSA) #9950 (1 µM, 4 hr) using Acetyl-Histone H4 (Lys12) (D2W60) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red).

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 H4 (Lys12) (D2W6O) 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, 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 HeLa cells, untreated (blue) or treated with SAHA (green) using Acetyl-Histone H4 (Lys12) (D2W6O) Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 fragment (Alexa Fluor® 488 Conjugate) #4412 was used as 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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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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