WB
Western blot analysis of DNA PKcs on different lysates with Rabbit anti-DNA PKcs antibody at 1/5,000 dilution. Lane 1: K-562 cell lysate Lane 2: NIH/3T3 cell lysate Lane 3: PC-12 cell lysate Lysates/proteins at 15 µg/Lane. Predicted band size: 469 kDa Observed band size: 469 kDa Exposure time: 5 seconds; ECL: K1801; 3-8% 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 at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-DNA PKcs antibody at 1/200 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 at 1/200 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.ICC/IF
Immunocytochemistry analysis of K-562 cells labeling DNA PKcs with Rabbit anti-DNA PKcs antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-DNA PKcs antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) 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. Beta tubulin ( red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.IP
DNA PKcs was immunoprecipitated from 0.2 mg NIH/3T3 cell lysate with Rabbit anti-DNA PKcs antibody at 2µg/10µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-DNA PKcs antibody at 1/1,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: NIH/3T3 cell lysate (input), Lane 2:Rabbit anti-DNA PKcs antibody IP in NIH/3T3 cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-DNA PKcs antibody in NIH/3T3 cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 6 seconds.| Product Name | DNA PKcs Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide within Human DNA PKcs aa 4062-4109 / 4128. |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ICC/IFIHCIPWB |
| WB:1:5000 IHC:1:50-1:200 ICC/IF:1:50-1:200 IP:1-2μg/sample |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | PRKDC |
|---|---|
| Gene Synonyms | HYRC p350 DNAPK DNPK1 HYRC1 IMD26 XRCC7 DNAPKc DNA-PKC DNA-PKcs |
| Gene Full Name | protein kinase, DNA-activated, catalytic subunit |
| Gene Summary | This gene encodes the catalytic subunit of the DNA-dependent protein kinase (DNA-PK). It functions with the Ku70/Ku80 heterodimer protein in DNA double strand break repair and recombination. The protein encoded is a member of the PI3/PI4-kinase family.[provided by RefSeq, Jul 2010] |
| Molecular Weight(MW) | 469kDa |
| Cellular Localization | Nucleus, nucleolus. |
WB
Western blot analysis of DNA PKcs on different lysates with Rabbit anti-DNA PKcs antibody at 1/5,000 dilution. Lane 1: K-562 cell lysate Lane 2: NIH/3T3 cell lysate Lane 3: PC-12 cell lysate Lysates/proteins at 15 µg/Lane. Predicted band size: 469 kDa Observed band size: 469 kDa Exposure time: 5 seconds; ECL: K1801; 3-8% 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 at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-DNA PKcs antibody at 1/200 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 at 1/200 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.
ICC/IF
Immunocytochemistry analysis of K-562 cells labeling DNA PKcs with Rabbit anti-DNA PKcs antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-DNA PKcs antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) 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. Beta tubulin ( red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
IP
DNA PKcs was immunoprecipitated from 0.2 mg NIH/3T3 cell lysate with Rabbit anti-DNA PKcs antibody at 2µg/10µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-DNA PKcs antibody at 1/1,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: NIH/3T3 cell lysate (input), Lane 2:Rabbit anti-DNA PKcs antibody IP in NIH/3T3 cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-DNA PKcs antibody in NIH/3T3 cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 6 seconds.| Application Notes | WB:1:5000 IHC:1:50-1:200 ICC/IF:1:50-1:200 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | 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. |
Data sheet for OM642889