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8346
Acetyl-Histone H4 Antibody Sampler Kit

Acetyl-Histone H4 Antibody Sampler Kit #8346

 Image 1

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.

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 Image 2

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.

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 Image 3

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

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Western Blotting Image 4

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

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Western Blotting Image 5

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.

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Western Blotting Image 6

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).

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Western Blotting Image 7

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

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Western Blotting Image 8

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.

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Western Blotting Image 9

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).

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Flow Cytometry Image 10

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.

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IHC-P (paraffin) Image 11

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).

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IHC-P (paraffin) Image 12

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).

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IF-IC Image 13

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).

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IHC-P (paraffin) Image 14

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

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Chromatin IP Image 15

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Acetyl-Histone H4 (Lys12) (D2W6O) Rabbit mAb or 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.

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IHC-P (paraffin) Image 16

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

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Chromatin IP Image 17

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Acetyl-Histone H4 (Lys5) (D12B3) Rabbit mAb or 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.

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Acetyl-Histone H4 (Lys5) (D12B3) Rabbit mAb 8647 20 µl
  • WB
  • IP
  • IHC
  • ChIP
H M R Mk 11 Rabbit IgG
Acetyl-Histone H4 (Lys8) Antibody 2594 20 µl
  • WB
H M R Mk 11 Rabbit 
Acetyl-Histone H4 (Lys12) (D2W6O) Rabbit mAb 13944 20 µl
  • WB
  • IP
  • IF
  • F
  • ChIP
H M R Mk 11 Rabbit IgG
Histone H4 (L64C1) Mouse mAb 2935 20 µl
  • WB
  • IHC
H M R Mk 11 Mouse IgG1
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 
Anti-mouse IgG, HRP-linked Antibody 7076 100 µl
  • WB
Horse 

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 two western blots with each antibody.

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

Polyclonal antibodies are produced by immunizing rabbits with synthetic acetylated peptide corresponding to residue 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 residues surrounding acetylated Lys5 of human histone H4 protein, acetylated Lys12 of human histone H4 protein or amino terminal sequence of human histone H4.

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).

  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
Entrez-Gene Id
8359
Swiss-Prot Acc.
P62805
For Research Use Only. Not For Use In Diagnostic Procedures.

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