Histone H4 (acetyl K16) Recombinant Rabbit Monoclonal Antibody [JB21-44]
cat.: ET7107-89
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P, FC, IF-Cell, IF-Tissue, ChIP
Clonality: Monoclonal
Clone number: JB21-44
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: 11 kDa
Isotype: IgG
Immunogen: Recombinant protein within Human Histone H4 (acetyl K16) aa 1-50 / 103.
Positive control: HeLa cell lysate, HeLa treated with 1μM TSA for 18 hours 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 colon tissue, mouse colon tissue.
Subcellular location: Chromosome, Nucleosome core, Nucleus.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
  ChIP
1:1,000
1:50-1:200
1:50-1:200
1:200-1:1,000
1:1,000
Use 0.5~2 μg for 25 μg of chromatin.
Uniprot #: SwissProt: P62805 Human | P62806 Mouse | P62804 Rat
Alternative names: Histone gene cluster 1, H4A Histone gene cluster 2, H4 dJ160A22.1 dJ160A22.2 dJ221C16.1 dJ221C16.9 FO108 H4 H4 histone family, member A H4 histone family, member B H4 histone family, member C H4 histone family, member D H4 histone family, member E H4 histone family, member G H4 histone family, member H H4 histone family, member I H4 histone family, member J H4 histone family, member K H4 histone family, member M H4 histone family, member N H4 histone, family 2 H4/A H4/B H4/C H4/D H4/E H4/G H4/H H4/I H4/J H4/K H4/M H4/N H4/O H4/p H4_HUMAN H4F2 H4F2iii H4F2iv H4FA H4FB H4FC H4FD H4FE H4FG H4FH H4FI H4FJ H4FK HIST1 cluster, H4A HIST1 cluster, H4B HIST1 cluster, H4D HIST2H4 Hist4 cluster, H4 Hist4h4 histone 1, H4a histone 1, H4c histone 1, H4d histone 1, H4f histone 1, H4h histone 1, H4i histone 1, H4j histone 1, H4k histone 1, H4l histone 2, H4a ......
Images
ET7107-89_1.jpg Fig1: Western blot analysis of Histone H4 (acetyl K16) on different lysates with Rabbit anti-Histone H4 (acetyl K16) antibody (ET7107-89) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: HeLa treated with 1μM TSA for 18 hours cell lysate
Lane 3: C6 cell lysate
Lane 4: C6 treated with 1μM TSA for 18 hours cell lysate
Lane 5: NIH/3T3 cell lysate
Lane 6: NIH/3T3 treated with 400nM TSA for 18 hours cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 11 kDa
Observed band size: 11 kDa

Exposure time: 3 minutes; 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 (ET7107-89) 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.
ET7107-89_2.jpg Fig2: Immunocytochemistry analysis of HeLa cells labeling Histone H4 (acetyl K16) with Rabbit anti-Histone H4 (acetyl K16) antibody (ET7107-89) at 1/100 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 H4 (acetyl K16) antibody (ET7107-89) at 1/100 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.
ET7107-89_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-Histone H4 (acetyl K16) antibody (ET7107-89) 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 (ET7107-89) 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.
ET7107-89_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-Histone H4 (acetyl K16) antibody (ET7107-89) 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 (ET7107-89) 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.
ET7107-89_5.jpg Fig5: Flow cytometric analysis of HeLa cells labeling Histone H4 (acetyl K16).

Cells were fixed and permeabilized. Then stained with the primary antibody (ET7107-89, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
ET7107-89_6.jpg Fig6: Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Histone H4 (acetyl K16) (ET7107-89) 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.