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9927
Acetyl-Histone H3 Antibody Sampler Kit

Acetyl-Histone H3 Antibody Sampler Kit #9927

Western Blotting Image 1

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

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Western Blotting Image 2

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.

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Chromatin IP-seq Image 3

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across GAPDH, a known target gene of H3K9Ac (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.

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Western Blotting Image 4

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).

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Chromatin IP-seq Image 5

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and Acetyl-Histone H3 (Lys14) (D4B9) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across VMP1 gene. For additional ChIP-seq tracks, please download the product data sheet.

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Chromatin IP-seq Image 6

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and Acetyl-Histone H3 (Lys18) (D8Z5H) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across NPR3 gene. For additional ChIP-seq tracks, please download the product data sheet.

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Western Blotting Image 7

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).

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Chromatin IP-seq Image 8

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and Acetyl-Histone H3 (Lys27) (D5E4) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across GAPDH, a known target gene of H3K27Ac (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.

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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 H3 (Lys27) (D5E4) XP® Rabbit mAb (upper) or Histone H3 (D1H2) XP® Rabbit mAb #4499 (lower).

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Western Blotting Image 10

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).

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Western Blotting Image 11

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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IHC-P (paraffin) Image 12

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

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Chromatin IP Image 13

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Acetyl-Histone H3 (Lys9) (C5B11) 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 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.

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IHC-P (paraffin) Image 14

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).

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Chromatin IP Image 15

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Acetyl-Histone H3 (Lys14) (D4B9) 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 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.

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Flow Cytometry Image 16

Flow cytometric analysis of HeLa cells, untreated (blue) or treated with Trichostatin A #9950 (1 μM, 4 hours; green) using Acetyl-Histone H3 (Lys14) (D4B9) Rabbit mAb #7627 (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

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Chromatin IP Image 17

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Acetyl-Histone H3 (Lys18) (D8Z5H) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. 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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IHC-P (paraffin) Image 18

Immunohistochemical analysis of paraffin-embedded transitional epithelial carcinoma of the bladder using Acetyl-Histone H3 (Lys18) (D8Z5H) Rabbit mAb.

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Flow Cytometry Image 19

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.

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Chromatin IP Image 20

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Acetyl-Histone H3 (Lys27) (D5E4) XP® 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 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.

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Flow Cytometry Image 21

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.

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Flow Cytometry Image 22

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).

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IF-IC Image 23

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).

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IHC-P (paraffin) Image 24

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

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IF-IC Image 25

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).

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IF-IC Image 26

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).

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IF-IC Image 27

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).

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IHC-P (paraffin) Image 28

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).

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Flow Cytometry Image 29

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.

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Histone H3 (D1H2) XP® Rabbit mAb 4499 20 µl
  • WB
  • IHC
  • IF
  • F
H M R Mk 17 Rabbit IgG
Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb 9649 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
  • ChIP
H M R Mk Z 17 Rabbit IgG
Acetyl-Histone H3 (Lys14) (D4B9) Rabbit mAb 7627 20 µl
  • WB
  • IP
  • IF
  • F
  • ChIP
H M R Mk 17 Rabbit IgG
Acetyl-Histone H3 (Lys18) (D8Z5H) Rabbit mAb 13998 20 µl
  • WB
  • IP
  • IHC
  • F
  • ChIP
H M R Mk Sc 17 Rabbit IgG
Acetyl-Histone H3 (Lys27) (D5E4) XP® Rabbit mAb 8173 20 µl
  • WB
  • IF
  • F
  • ChIP
H M R Mk 17 Rabbit IgG
Acetyl-Histone H3 (Lys56) Antibody 4243 20 µl
  • WB
H M R Mk 17 Rabbit 
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

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

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

Polyclonal antibodies are produced by immunizing rabbits with synthetic acetylated peptides corresponding to residues surrounding Lys56 of human Histone H3. 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 Lys18 of human histone H3 protein, Lys14 of human histone H3 protein, acetylated Lys27 of human Histone H3 protein, or the amino terminus of histone H3 in which Lys9 is acetylated.

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
8350
Swiss-Prot Acc.
P68431
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

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