Histone H3 (acetyl K56) Recombinant Rabbit Monoclonal Antibody [SU30-10]
cat.: ET1608-9
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P, ChIP, CUT&Tag-seq
Clonality: Monoclonal
Clone number: SU30-10
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 15 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human Histone H3 aa 31-80 / 136.
Positive control: HeLa cell lysate, C6 cell lysate, C6 treated with 1μM TSA for 18 hours cell lysate, NIH/3T3 cell lysate, NIH/3T3 treated with 400nM TSA for 18 hours cell lysate, HeLa, human kidney tissue, rat kidney tissue, human skin tissue, human colon carcinoma tissue, human breast carcinoma tissue, human esophageal carcinoma tissue.
Subcellular location: Nucleus, Chromosome, Nucleosome core.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  ChIP
1:500-1:2,000
1:50-1:200
1:50-1:200
1:200-1:5,000
Use 0.5~2 μg for 25 μg of chromatin.
Uniprot #: SwissProt: P68431 Human | P84243 Human | Q71DI3 Human | P68433 Mouse | Q6LED0 Rat
Alternative names: H3.3A HIST1 cluster, H3E H3 histone family, member A H3.1 H3/l H3F3 H3FF H3FJ H3FL Histone gene cluster 1, H3 histone family, member E histone H3.1t Histone H3/o FLJ92264 H 3 H3 H3 histone family, member B H3 histone family, member C H3 histone family, member D H3 histone family, member F H3 histone family, member H H3 histone family, member I H3 histone family, member J H3 histone family, member K H3 histone family, member L H3 histone family, member T H3 histone, family 3A H3/A H3/b H3/c H3/d h3/f H3/h H3/i H3/j H3/k H3/t H31_HUMAN H3F1K H3F3A H3FA H3FB H3FC H3FD H3FH H3FI H3FK HIST1 cluster, H3A HIST1 cluster, H3B HIST1 cluster, H3C HIST1 cluster, H3D HIST1 cluster, H3F HIST1 cluster, H3G HIST1 cluster, H3H HIST1 cluster, H3I HIST1 cluster, H3J HIST1H3A HIST1H3B HIST1H3C HIST1H3D HIST1H3E HIST1H3F HIST1H3G HIST1H3H HIST1H3I HIST1H3J HIST3H3 Histone 1, H......
Images
ET1608-9_1.jpg Fig1: Western blot analysis of Histone H3 (acetyl K56) on different lysates with Rabbit anti-Histone H3 (acetyl K56) antibody (ET1608-9) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: C6 cell lysate
Lane 3: C6 treated with 1μM TSA for 18 hours cell lysate
Lane 4: NIH/3T3 cell lysate
Lane 5: NIH/3T3 treated with 400nM TSA for 18 hours cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 15 kDa
Observed band size: 15 kDa

Exposure time: 14 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1608-9) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1608-9_2.jpg Fig2: Immunocytochemistry analysis of HeLa cells labeling Histone H3 (acetyl K56) with Rabbit anti-Histone H3 (acetyl K56) antibody (ET1608-9) at 1/250 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Histone H3 (acetyl K56) antibody (ET1608-9) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ET1608-9_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Histone H3 (acetyl K56) antibody (ET1608-9) at 1/5,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-9) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1608-9_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-Histone H3 (acetyl K56) antibody (ET1608-9) at 1/5,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-9) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1608-9_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human skin tissue with Rabbit anti-Histone H3 (acetyl K56) antibody (ET1608-9) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-9) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1608-9_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-Histone H3 (acetyl K56) antibody (ET1608-9) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-9) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1608-9_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-Histone H3 (acetyl K56) antibody (ET1608-9) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-9) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1608-9_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded human esophageal carcinoma tissue with Rabbit anti-Histone H3 (acetyl K56) antibody (ET1608-9) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-9) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1608-9_9.jpg Fig9: Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Histone H3 (acetyl K56) (ET1608-9) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. 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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