Anti-DNA PKcs (phospho S2056) antibody - ChIP Grade

• Supplier: Abcam
• Catalog: ab18192
• Prices: $404.00
• Sizes: 100 µg
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Applications: FC IHC-P WB ChIP ICC/IF ICC/IF
• Species Reactivities: Human, Mouse
• Antigen: Synthetic peptide conjugated to KLH derived from within residues 2050 - 2150 of Human DNA PKcs
• Blocking Peptide: Human DNA PKcs (phospho S2056) peptide
• Description: Rabbit Polyclonal
• Gene: PRKDC
• Conjugate: Unconjugated

Product images

All lanes : Anti-DNA PKcs (phospho S2056) antibody - ChIP Grade (ab18192) at 1 µg/mlLane 1 : HeLa Gamma Irradiated Whole Cell Lysate Pack (ab13823) at 20 µgLane 2 : 20ug of untreated HeLa cell extractLane 3 : HeLa Gamma Irradiated Whole Cell Lysate Pack (ab13823) at 20 µg with Human DNA PKcs (phospho S2056) peptide (ab20406) at 1 µg/mlLane 4 : 20ug of untreated HeLa cell extract with Human DNA PKcs (phospho S2056) peptide (ab20406) at 1 µg/mlLane 5 : HeLa Gamma Irradiated Whole Cell Lysate Pack (ab13823) at 20 µg with Human DNA PKcs peptide (ab20407) at 1 µg/mlLane 6 : 20ug of untreated HeLa cell extract with Human DNA PKcs peptide (ab20407) at 1 µg/mlSecondaryAlexa Fluor Goat polyclonal to Rabbit IgG (700) at 1/10000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.

ICC/IF image of ab18192 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal got serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab18192, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min)HeLa, Hek293 and MCF7 cells at 1µg/ml, and in 100% methanol fixed (5 min) HeLa, Hek293 and HepG2 cells at 5µg/ml.

All lanes : Anti-DNA PKcs (phospho S2056) antibody - ChIP Grade (ab18192) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 2 : Hela Whole Cell Lysate - Bleomycin Treated (20U/ml) Lane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with Human DNA PKcs (phospho S2056) peptide (ab20406) at 1 µg/mlLane 4 : Hela Whole Cell Lysate - Bleomycin Treated (20U/ml) with Human DNA PKcs (phospho S2056) peptide (ab20406) at 1 µg/mlLane 5 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with Human DNA PKcs peptide (ab20407) at 1 µg/mlLane 6 : Hela Whole Cell Lysate - Bleomycin Treated (20U/ml) with Human DNA PKcs peptide (ab20407) at 1 µg/mlLysates/proteins at 20 µg per lane.SecondaryGoat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilutionPerformed under reducing conditions.

Overlay histogram showing Jurkat cells stained with ab18192 (blue line). The cells were prepared as follows: Spin down, wash in FACS buffer 1x, fix, wash 2x, and stain with primary antibody overnight. Buffer was was 0.1% sodium azide with FBS in PBS. Cells fixed with paraformaldehyde and then permeabilized with Triton X-100 and NP40. Cell population was gated by isolating cell population from plot of SSC-A / FSA-A. The cells were then incubated the antibody at 1/100 for 24 hours. The secondary antibody used was FITC-conjugated Goat anti-Rabbit polyclonal at 1/100 dilution. Isotype control antibody (red line) was used under the same conditions.See Abreview

ab18192 staining DNA PKcs in Human aorta tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 100% blocking agent for 20 minutes; antigen retrieval was enzymatic using Proteinase K. Samples were incubated with primary antibody (1/350 in diluent) for 15 hours at 4°C. A biotinylated Goat anti-rabbit polyclonal (1/800) was used as the secondary antibody.See Abreview

ab18192 staining DNA PKcs (phospho S2056) in leukaemia cell lines by Immunocytochemistry/ Immunofluorescence. Cells were either untreated (control) or treated with ionizing radiation (IR) or ultrasound (US). Following treatment, cells were fixed with paraformaldehyde and permabilized/blocked with 2% BSA/0.05% Triton X-100/Tris-buffered saline. Samples were incubated with primary antibody (1/600 in diluent) for 2 hours. An AlexaFluor®555-conjugated anti-rabbit IgG (1/400) was used as the secondary antibody.

Product References

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