Histone H3 (acetyl K27) Rabbit Polyclonal Antibody
cat.: HA500046
Product Type: Rabbit polyclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P, IF-Cell, ChIP
Clonality: Polyclonal
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage buffer: 1*TBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Immunogen affinity purified.
Molecular weight: Predicted band size: 15 kDa
Isotype: IgG
Immunogen: Synthetic peptide within human Histone H3 aa 20-40 (acetyl K27).
Positive control: HeLa (Human cervical adenocarcinoma cell) cell lysate, HeLa treated with 500ng/mL TSA for 4 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, human skin tissue, human colon tissue, mouse colon tissue, rat colon tissue.
Subcellular location: Chromosome, Nucleosome core, Nucleus.
Recommended Dilutions:
  WB
  IHC-P
  IF-Cell
  ChIP
1:50,000
1:100-1:500
1:2,000-1:5,000
Use 0.5~2 μg for 25 μg of chromatin.
Uniprot #: SwissProt: P68431 Human | P84243 Human | Q16695 Human | Q71DI3 Human | Q6NXT2 Human
Alternative names: H3 histone family member E pseudogene H3 histone family, member A H3/A H31_HUMAN H3F3 H3FA Hist1h3a HIST1H3B HIST1H3C HIST1H3D HIST1H3E HIST1H3F HIST1H3G HIST1H3H HIST1H3I HIST1H3J HIST3H3 histone 1, H3a Histone cluster 1, H3a Histone H3 3 pseudogene Histone H3.1 Histone H3/a Histone H3/b Histone H3/c Histone H3/d Histone H3/f Histone H3/h Histone H3/i Histone H3/j Histone H3/k Histone H3/l H3K27ac
Images
HA500046_1.jpg Fig1: Western blot analysis of Histone H3 (acetyl K27) on different lysates with Rabbit anti-Histone H3 (acetyl K27) antibody (HA500046) at 1/50,000 dilution.

Lane 1: HeLa (Human cervical adenocarcinoma cell) cell lysate
Lane 2: HeLa treated with 500ng/mL TSA for 4 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 15 µg/Lane.
Exposure time: 59 seconds; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA500046, 1/50,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 15 kDa
Observed band size: 15 kDa
HA500046_2.jpg Fig2: Application: Immunohistochemistry (IHC-P)

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

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

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: HA500046, 1/500, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA500046_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human skin tissue with Rabbit anti-Histone H3 (acetyl K27) antibody (HA500046) at 1/500 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 (HA500046) at 1/500 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.
HA500046_4.jpg Fig4: 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℃.

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: HA500046, 1/500, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA500046_5.jpg Fig5: 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℃.

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: HA500046, 1/500, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA500046_6.jpg Fig6: Application: Immunocytochemistry (IF-cell)

Species: Human
Sample: HeLa (Human cervix adenocarcinoma epithelial cell) treated with 500ng/mL TSA for 4 hours

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: HA500046, 1/5,000, overnight at 4℃.
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).
HA500046_7.jpg Fig7: Application: Immunocytochemistry (IF-cell)

Species: Mouse
Sample: NIH/3T3 (Mouse fibroblast) treated with 400nM TSA for 18 hours

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: HA500046, 1/2,500, overnight at 4℃.
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).
HA500046_8.jpg Fig8: Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells treated with 500ng/mL TSA for 4 hours with Histone H3 (acetyl K27) (HA500046) 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”.