Histone H3 Recombinant Rabbit Monoclonal Antibody [JJ090-07]
cat.: ET1701-64
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P, ChIP, IP
Clonality: Monoclonal
Clone number: JJ090-07
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: Recombinant protein within human Histone H3 aa 85-136/136.
Positive control: HeLa cell lysate, A549 cell lysate, HT-29 cell lysate, HEK-293 cell lysate, C2C12 cell lysate, L-929 cell lysate, C6 cell lysate, HeLa, human kidney tissue, human skin tissue, mouse liver tissue, mouse kidney tissue, rat liver tissue, rat kidney tissue, rat skin tissue, human liver tissue, mouse testis tissue, rat testis tissue.
Subcellular location: Nucleus, Chromosome.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  ChIP
  IP
1:20,000
1:100
1:100
1:5,000-1:10,000
Use 0.5~2 μg for 25 μg of chromatin.
1-2μg/sample
Uniprot #: SwissProt: P68431 Human | P84243 Human | Q16695 Human | Q6NXT2 Human | Q71DI3 Human | P68433 Mouse | P84228 Mouse | Q6LED0 Rat
Alternative names: H3 histone family, member A H3/A H31_HUMAN H3FA Hist1h3a HIST1H3B HIST1H3C HIST1H3D HIST1H3E HIST1H3F HIST1H3G HIST1H3H HIST1H3I HIST1H3J histone 1, H3a Histone cluster 1, H3a 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
Images
ET1701-64_1.jpg Fig1: Western blot analysis of Histone H3 on different lysates with Rabbit anti-Histone H3 antibody (ET1701-64) at 1/20,000 dilution and competitor's antibody at 1/5,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: A549 cell lysate
Lane 3: HT-29 cell lysate
Lane 4: HEK-293 cell lysate
Lane 5: C2C12 cell lysate
Lane 6: L-929 cell lysate
Lane 7: C6 cell lysate

Lysates/proteins at 10 µg/Lane.

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

Exposure time: 18 seconds;

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 (ET1701-64) at 1/20,000 dilution and competitor's antibody at 1/5,000 dilution were 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.
ET1701-64_2.jpg Fig2: Immunocytochemistry analysis of HeLa cells labeling Histone H3 with Rabbit anti-Histone H3 antibody (ET1701-64) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Histone H3 antibody (ET1701-64) at 1/100 dilution in 2% negative goat serum 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.
ET1701-64_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Histone H3 antibody (ET1701-64) at 1/10,000 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 (ET1701-64) at 1/10,000 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.
ET1701-64_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human skin tissue with Rabbit anti-Histone H3 antibody (ET1701-64) 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 (ET1701-64) 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.
ET1701-64_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-Histone H3 antibody (ET1701-64) at 1/10,000 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 (ET1701-64) at 1/10,000 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.
ET1701-64_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-Histone H3 antibody (ET1701-64) at 1/10,000 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 (ET1701-64) at 1/10,000 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.
ET1701-64_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-Histone H3 antibody (ET1701-64) at 1/10,000 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 (ET1701-64) at 1/10,000 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.
ET1701-64_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-Histone H3 antibody (ET1701-64) at 1/10,000 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 (ET1701-64) at 1/10,000 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.
ET1701-64_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded rat skin tissue with Rabbit anti-Histone H3 antibody (ET1701-64) 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 (ET1701-64) 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.
ET1701-64_10.jpg Fig10: Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and Histone H3 (ET1701-64) / 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.
ET1701-64_11.jpg Fig11: Histone H3 was immunoprecipitated in 0.2mg HeLa cell lysate with ET1701-64 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using ET1701-64 at 1/20,000 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: ET1701-64 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of ET1701-64 in HeLa cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 1 minute 59 seconds
ET1701-64_12.jpg Fig12: Immunofluorescence analysis of paraffin-embedded human liver tissue labeling Histone H3 with Rabbit anti-Histone H3 antibody (ET1701-64) at 1/100 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1701-64, green) at 1/100 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
ET1701-64_13.jpg Fig13: Immunofluorescence analysis of paraffin-embedded mouse testis tissue labeling Histone H3 with Rabbit anti-Histone H3 antibody (ET1701-64) at 1/100 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1701-64, green) at 1/100 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
ET1701-64_14.jpg Fig14: Immunofluorescence analysis of paraffin-embedded rat testis tissue labeling Histone H3 with Rabbit anti-Histone H3 antibody (ET1701-64) at 1/100 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1701-64, green) at 1/100 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
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