Phospho-Histone H2A.X (S139) Recombinant Rabbit Monoclonal Antibody [SR33-09]
cat.: ET1602-2
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IHC-P |
| Clonality: | Monoclonal |
| Clone number: | SR33-09 |
| 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 phospho-peptide corresponding to residues surrounding Ser139 of Human Histone H2AX. |
| Positive control: | HeLa treated with 20μM Etoposide for 2 hours whole cell lysate, HeLa treated with UV for 2 hours whole cell lysate, NIH/3T3 treated with 25μM Etoposide for 5 hours whole cell lysate, HepG2 cell lysate–treated with etoposide, rat brain tissue, mouse testis tissue, mouse brain tissue, mouse small intestine tissue, human colon carcinoma tissue. |
| Subcellular location: | Nucleus, Chromosome |
| Recommended Dilutions:
WB IHC-P |
1:1,000-1:2,000 1:50-1:5000 |
| Uniprot #: | SwissProt: P16104 Human | P27661 Mouse Unigene: 2850 Rat |
| Alternative names: | AW228881 H2A histone family member X H2A.FX H2A.X H2a/x H2AFX H2AX H2AX histone H2AX_HUMAN Hist5.2ax Histone 2A Histone 2AX Histone H2A.X Histone H2AX RGD1566119 gamma H2A.X γ H2A.X γ-H2A.X |
Images
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Fig1:
Western blot analysis of Phospho-Histone H2A.X (S139) on different lysates with Rabbit anti-Phospho-Histone H2A.X (S139) antibody (ET1602-2) at 1/2,000 dilution. Lane 1: HeLa whole cell lysate Lane 2: HeLa treated with 20μM Etoposide for 2 hours whole cell lysate Lane 3: HeLa whole cell lysate Lane 4: HeLa treated with UV for 2 hours whole cell lysate Lane 5: NIH/3T3 whole cell lysate Lane 6: NIH/3T3 treated with 25μM Etoposide for 5 hours whole cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 15 kDa Observed band size: 15/20 kDa Exposure time: 53 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 (ET1602-2) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Western blot analysis of Phospho-Histone H2A.X (S139) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1602-2, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature. Positive control: Lane 1: HepG2 cell lysate–treated with etoposide Lane 2: HepG2 cell lysate–untreated |
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Fig3:
Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-Phospho-Histone H2A.X (S139) antibody (ET1602-2) at 1/500 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 (ET1602-2) 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. |
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Fig4:
Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-Phospho-Histone H2A.X (S139) antibody (ET1602-2) at 1/500 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 (ET1602-2) 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. |
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Fig5:
Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-Phospho-Histone H2A.X (S139) antibody (ET1602-2) at 1/50 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 (ET1602-2) at 1/50 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. |
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Fig6:
Immunohistochemical analysis of paraffin-embedded mouse small intestine tissue with Rabbit anti-Phospho-Histone H2A.X (S139) antibody (ET1602-2) at 1/500 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 (ET1602-2) 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. |
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Fig7:
Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-Phospho-Histone H2A.X (S139) antibody (ET1602-2) at 1/400 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 (ET1602-2) at 1/400 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. |
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Fig8:
Western blot analysis of Phospho-Histone H2A.X (S139) on different lysates with Rabbit anti-Phospho-Histone H2A.X (S139) antibody (ET1602-2) at 1/1,000 dilution. Lane 1: C6 treated with 25μM Etoposide for 5 hours whole cell lysate Lane 2: C6 whole cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 15 kDa Observed band size: 15 kDa Exposure time: 20 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 (ET1602-2) 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. |
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Fig9:
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-Phospho-Histone H2A.X (S139) antibody (ET1602-2) at 1/500 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 (ET1602-2) 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. |
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Fig10:
Immunohistochemical analysis of paraffin-embedded mouse breast tissue with Rabbit anti-Phospho-Histone H2A.X (S139) antibody (ET1602-2) 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 (ET1602-2) 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. |
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Fig11:
Immunohistochemical analysis of paraffin-embedded mouse skin tissue with Rabbit anti-Phospho-Histone H2A.X (S139) antibody (ET1602-2) at 1/500 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 (ET1602-2) 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. |
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