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9933
Acetyl-Histone Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Acetyl-Histone Antibody Sampler Kit #9933

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Acetyl-Histone Antibody Sampler Kit: Image 1

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).

Acetyl-Histone Antibody Sampler Kit: Image 2

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Acetyl-Histone H2B (Lys5) (D5H1S) 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 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.

Acetyl-Histone Antibody Sampler Kit: Image 3

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

Acetyl-Histone Antibody Sampler Kit: Image 4

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Histone H2A (D6O3A) 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 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.

Acetyl-Histone Antibody Sampler Kit: Image 5

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Histone H2B (D2H6) 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 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.

Acetyl-Histone Antibody Sampler Kit: Image 6

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).

Acetyl-Histone Antibody Sampler Kit: Image 7

CUT&RUN was performed with HeLa cells and Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb, using CUT&RUN Assay Kit #86652. 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 CUT&RUN-qPCR data).

Acetyl-Histone Antibody Sampler Kit: Image 8

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.

Acetyl-Histone Antibody Sampler Kit: Image 9

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.

Acetyl-Histone Antibody Sampler Kit: Image 10

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

Acetyl-Histone Antibody Sampler Kit: 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.

Acetyl-Histone Antibody Sampler Kit: Image 12

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.

Acetyl-Histone Antibody Sampler Kit: Image 13

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

Acetyl-Histone Antibody Sampler Kit: Image 14

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

Acetyl-Histone Antibody Sampler Kit: Image 15

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 chromosome 12 (upper), including GAPDH (lower), a known target gene of H3K9Ac (see additional figure containing ChIP-qPCR data).

Acetyl-Histone Antibody Sampler Kit: Image 16

CUT&RUN was performed with HeLa cells and Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figures show binding across chromosome 12 (upper), including GAPDH (lower), a known target gene of H3K9Ac (see additional figure containing CUT&RUN-qPCR data).

Acetyl-Histone Antibody Sampler Kit: Image 17

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).

Acetyl-Histone Antibody Sampler Kit: Image 18

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

Acetyl-Histone Antibody Sampler Kit: Image 19

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

Acetyl-Histone Antibody Sampler Kit: Image 20

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

Acetyl-Histone Antibody Sampler Kit: Image 21

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.

Acetyl-Histone Antibody Sampler Kit: Image 22

CUT&RUN was performed with HeLa cells and either Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human RPL30 Exon 3 Primers #7014, SimpleChIP® Human GAPDH Exon 1 Primers #5516, 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.

Acetyl-Histone Antibody Sampler Kit: Image 23

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

Acetyl-Histone Antibody Sampler Kit: Image 24

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

Acetyl-Histone Antibody Sampler Kit: Image 25

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

Acetyl-Histone Antibody Sampler Kit: Image 26

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).

Acetyl-Histone Antibody Sampler Kit: Image 27

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

Acetyl-Histone Antibody Sampler Kit: Image 28

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

Acetyl-Histone Antibody Sampler Kit: Image 29

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).

Acetyl-Histone Antibody Sampler Kit: Image 30

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

Acetyl-Histone Antibody Sampler Kit: Image 31

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).

Acetyl-Histone Antibody Sampler Kit: Image 32

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.

Acetyl-Histone Antibody Sampler Kit: Image 33

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

Acetyl-Histone Antibody Sampler Kit: Image 34

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).

Acetyl-Histone Antibody Sampler Kit: Image 35

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).

Acetyl-Histone Antibody Sampler Kit: Image 36

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).

Acetyl-Histone Antibody Sampler Kit: Image 37

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).

Acetyl-Histone Antibody Sampler Kit: Image 38

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.

To Purchase # 9933T
Product # Size Price
9933T
1 Kit  (8 x 20 µl) $ 554

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Acetyl-Histone H2B (Lys5) (D5H1S) XP® Rabbit mAb 12799 20 µl
  • WB
  • IP
  • IHC
  • IF
  • ChIP
H Mk M R 14 Rabbit IgG
Acetyl-Histone H2A (Lys5) Antibody 2576 20 µl
  • WB
  • IP
  • IHC
H Mk M R 14 Rabbit 
Histone H2A (D6O3A) Rabbit mAb 12349 20 µl
  • WB
  • IF
  • ChIP
GP H Mk M R Z 14 Rabbit IgG
Histone H2B (D2H6) Rabbit mAb 12364 20 µl
  • WB
  • IHC
  • ChIP
H Mk M R 14 Rabbit IgG
Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb 9649 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
  • ChIP
  • C&R
H Mk M R Z 17 Rabbit IgG
Histone H3 (D1H2) XP® Rabbit mAb 4499 20 µl
  • WB
  • IHC
  • IF
  • F
H Mk M R 17 Rabbit IgG
Acetyl-Histone H4 (Lys8) Antibody 2594 20 µl
  • WB
H Mk M R 11 Rabbit 
Histone H4 (D2X4V) Rabbit mAb 13919 20 µl
  • WB
  • IHC
  • IF
H Mk M R 11 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product 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 two Western blot experiments.

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.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide and are purified by protein A and peptide affinity chromatography. Monoclonal antibodies are produced by immunizing animals with recombinant human proteins or synthetic peptides.

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).

  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.

Limited Uses

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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.