Histone H4 (acetyl K12) Recombinant Rabbit Monoclonal Antibody [PSH25-67]
cat.: HA751986
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P, IP, ChIP
Clonality: Monoclonal
Clone number: PSH25-67
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: PBS (pH7.4).
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 11 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human Histone H4 aa 1-50 (acetyl K12).
Positive control: HeLa (Human cervical adenocarcinoma cell) cell lysate, HeLa treated with 1μM TSA for 24 hours cell lysate, NIH/3T3 (Mouse fibroblast) cell lysate, NIH/3T3 treated with 400nM TSA for 18 hours cell lysate, C6 (Rat glioma cell) cell lysate, C6 treated with 1μM TSA for 18 hours cell lysate.
Subcellular location: Nucleus. Chromosome.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  IP
  ChIP
1:5,000
1:2,000
1:500-1:1,000
1-2μg/sample
Use 0.5~2 μg for 25 μg of chromatin.
Uniprot #: SwissProt: P62805 Human | P62806 Mouse | P62804 Rat
Alternative names: dJ160A22.1 dJ160A22.2 dJ221C16.1 dJ221C16.9 FO108 H4 H4.k 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/p H4_HUMAN H4F2 H4F2iii H4F2iv H4FA H4FB H4FC H4FD H4FE H4FG H4FH H4FI H4FJ H4FK H4FM H4FN H4M HIST1H4A HIST1H4B HIST1H4C HIST1H4D HIST1H4E HIST1H4F HIST1H4H HIST1H4I HIST1H4J HIST1H4K HIST1H4L HIST2H4 HIST2H4A Hist4h4 Histone 1 H4a Histone 1 H4b Histone 1 H4c Histone 1 H4d Histone 1 H4e Histone 1 H4f Histone 1 H4h Histone 1 H4i Histone 1 H4j Histone 1 H4k Histone 1 H4l Histone 2 H4a histone 4 H4 Histone H4 MGC24116 H4K12ac
Images
HA751986_1.jpg Fig1: Western blot analysis of Histone H4 (acetyl K12) on different lysates with Rabbit anti-Histone H4 (acetyl K12) antibody (HA751986) at 1/50,000 dilution.

Lane 1: HeLa (Human cervical adenocarcinoma cell) cell lysate
Lane 2: HeLa treated with 1μM TSA for 24 hours cell lysate
Lane 3: NIH/3T3 (Mouse fibroblast) cell lysate
Lane 4: NIH/3T3 treated with 400nM TSA for 18 hours cell lysate
Lane 5: C6 (Rat glioma cell) cell lysate
Lane 6: C6 treated with 1μM TSA for 18 hours cell lysate

Lysates/proteins at 10 µg/Lane.
Exposure time: 2 seconds; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA751986, 1/50,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 11 kDa
Observed band size: 15 kDa
HA751986_2.jpg Fig2: Application: Immunocytochemistry (IF-cell)

Species: Human
Sample: HeLa (Human cervical adenocarcinoma cell)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.

Primary antibody: HA751986, 1/2,000, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).
HA751986_3.jpg Fig3: Application: Immunocytochemistry (IF-cell)

Species: Mouse
Sample: NIH/3T3 (Mouse fibroblast)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.

Primary antibody: HA751986, 1/2,000, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).
HA751986_4.jpg Fig4: Application: Immunocytochemistry (IF-cell)

Species: Rat
Sample: C6 (Rat glioma cell)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.

Primary antibody: HA751986, 1/2,000, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).
HA751986_5.jpg Fig5: Application: Immunohistochemistry (IHC-P)

Species: Human
Tissue: Brain
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA751986, 1/500, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA751986_6.jpg Fig6: Application: Immunohistochemistry (IHC-P)

Species: Mouse
Tissue: Colon
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA751986, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA751986_7.jpg Fig7: Application: Immunohistochemistry (IHC-P)

Species: Rat
Tissue: Colon
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA751986, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA751986_8.jpg Fig8: Immunoprecipitation (IP)

Histone H4 (acetyl K12) was immunoprecipitated in 0.2 mg HeLa (Human cervix adenocarcinoma epithelial cell) cell lysate with HA751986 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751986 at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: HeLa cell lysate (input)
Lane 2: HA751986 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of HA751986 in HeLa cell lysate

Exposure time: 2 seconds
Blocking: 5% NFDM/TBST, 1 hour at room temperature

Primary dilution: HA751986, 1/5,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature

Predicted band size: 11 kDa
Observed band size: 15 kDa
HA751986_9.jpg Fig9: Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells with Histone H4 (acetyl K12) (HA751986) / Competitor's antibody / 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”.