WB
Western blot analysis of various lysates using Phospho-Histone H2AX-S139 Rabbit mAb at 1:1000 dilution incubated overnight at 4℃. 293T and C6 cells were treated by UV at room temperature for 15-30 minutes. Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) at 1:10000 dilution. Lysates/proteins: 30 μg per lane. Blocking buffer: 3% nonfat dry milk in TBST. Detection: ECL Basic Kit. Exposure time: 20sWB
Western blot analysis of lysates from NIH/3T3 cells using Phospho-Histone H2AX-S139 Rabbit mAb at 1:1000 dilution incubated overnight at 4℃. NIH/3T3 cells were treated by UV at room temperature for 15-30 minutes. Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) at 1:10000 dilution. Lysates/proteins: 30 μg per lane. Blocking buffer: 3 % nonfat dry milk in TBST. Detection: ECL Basic Kit. Exposure time: 45s.ICC/IF
Confocal imaging of NIH/3T3 cells (treated with UV) and NIH/3T3 cells (untreated) using Phospho-Histone H2AX-S139 Rabbit mAb (dilution 1:2000) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.ICC/IF
Confocal imaging of NIH/3T3 cells (treated with UV) and NIH/3T3 cells (untreated) using Phospho-Histone H2AX-S139 Rabbit mAb (dilution 1:2000) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.ICC/IF
Confocal imaging of C6 cells (treated with UV) and C6 cells (untreated) using Phospho Histone H2AX-S139 Rabbit mAb (dilution 1:2000) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.IP
Immunoprecipitation of Phospho-Histone H2AX-S139 from 882 µg extracts of 293T cells treated with UV(90mJ/cm2，2 h) was performed using 1 µg of Phospho-Histone H2AX-S139 Rabbit mAb. Rabbit Control IgG was used to precipitate the Control IgG sample. IP samples were eluted with 1x non-reducing Laemmli Buffer. The Input lane represents 10% of the total input. Western blot analysis of immunoprecipitates was conducted using Phospho-Histone H2AX-S139 Rabbit mAb at a dilution of 1:1000.| Product Name | Phospho-Histone H2AX-S139 Rabbit mAb |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide. This information is considered to be commercially sensitive. |
| Modification | P-S139 |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ICC/IFIPWB |
| WB:1:1000-1:4000 ICC/IF:1:400-1:4000 IP:0.5μg-4μg antibody for 200μg-900μg extracts of whole cells |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Affinity purified |
| Gene Symbol | H2AX |
|---|---|
| Gene Synonyms | H2A.X H2A/X H2AFX |
| Gene Full Name | H2A.X variant histone |
| Gene Summary | Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Two molecules of each of the four core histones (H2A, H2B, H3, and H4) form an octamer, around which approximately 146 bp of DNA is wrapped in repeating units, called nucleosomes. The linker histone, H1, interacts with linker DNA between nucleosomes and functions in the compaction of chromatin into higher order structures. This gene encodes a replication-independent histone that is a member of the histone H2A family, and generates two transcripts through the use of the conserved stem-loop termination motif, and the polyA addition motif. [provided by RefSeq, Oct 2015] |
| Alternative Names | H2A.X H2A/X H2AFX |
| Molecular Weight(MW) | 15kDa(Observed band size:17kDa) |
| Cellular Localization | Chromosome, Nucleus. |
WB
Western blot analysis of various lysates using Phospho-Histone H2AX-S139 Rabbit mAb at 1:1000 dilution incubated overnight at 4℃. 293T and C6 cells were treated by UV at room temperature for 15-30 minutes. Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) at 1:10000 dilution. Lysates/proteins: 30 μg per lane. Blocking buffer: 3% nonfat dry milk in TBST. Detection: ECL Basic Kit. Exposure time: 20s
WB
Western blot analysis of lysates from NIH/3T3 cells using Phospho-Histone H2AX-S139 Rabbit mAb at 1:1000 dilution incubated overnight at 4℃. NIH/3T3 cells were treated by UV at room temperature for 15-30 minutes. Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) at 1:10000 dilution. Lysates/proteins: 30 μg per lane. Blocking buffer: 3 % nonfat dry milk in TBST. Detection: ECL Basic Kit. Exposure time: 45s.
ICC/IF
Confocal imaging of NIH/3T3 cells (treated with UV) and NIH/3T3 cells (untreated) using Phospho-Histone H2AX-S139 Rabbit mAb (dilution 1:2000) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.
ICC/IF
Confocal imaging of NIH/3T3 cells (treated with UV) and NIH/3T3 cells (untreated) using Phospho-Histone H2AX-S139 Rabbit mAb (dilution 1:2000) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.
ICC/IF
Confocal imaging of C6 cells (treated with UV) and C6 cells (untreated) using Phospho Histone H2AX-S139 Rabbit mAb (dilution 1:2000) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.
IP
Immunoprecipitation of Phospho-Histone H2AX-S139 from 882 µg extracts of 293T cells treated with UV(90mJ/cm2，2 h) was performed using 1 µg of Phospho-Histone H2AX-S139 Rabbit mAb. Rabbit Control IgG was used to precipitate the Control IgG sample. IP samples were eluted with 1x non-reducing Laemmli Buffer. The Input lane represents 10% of the total input. Western blot analysis of immunoprecipitates was conducted using Phospho-Histone H2AX-S139 Rabbit mAb at a dilution of 1:1000.| Application Notes | WB:1:1000-1:4000 ICC/IF:1:400-1:4000 IP:0.5μg-4μg antibody for 200μg-900μg extracts of whole cells |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at -20℃. Avoid freeze / thaw cycles. |
| Storage Buffer | PBS with 0.09% Sodium azide, 0.05% BSA, 50% glycerol, pH7.3. |
