![Western Blot: DNA-PKcs [p Thr2609] Antibody [NB600-897] - Lane 1: Fus1 untreated, Lane 2: Fus1 IR (20Gy, 4h), Lane 3: Fus1 DNAPK inhibitor + IR, Lane 4: MO59J (DNAPK-) untreated, Lane 5: MO59J IR, Lane 6: Fus1 untreated, Lane 7: Fus1 IR (20Gy, 4h), Lane 8: Fus1 DNAPK inhibitor + IR, Lane 9: MO59J untreated, Lane 10: MO59J IR. Lanes 1-5 are nuclear extract whereas lanes 6-10 are whole cell lysates. MO59J is a cell line that lacks DNA-PKcs. FUS1 is the matched cell line complemented with a chromosomal fragment containing the DNA-PKcs gene. Approximately 20 ug of lysate was run on SDS-PAGE and transferred onto nitrocellulose, followed by reaction with a 1:1,000 dilution of anti-DNAPKcs antibody. Detection occurred using a 1:5,000 dilution of HRP-labeled Goat anti-Rabbit IgG for 1 hour at room temperature. A chemiluminescence system was used for signal detection (Roche) using a 1 min exposure time.](http://images.novusbio.com/fullsize/DNA-PKcs-[p-Thr2609]-Antibody-Western-Blot-NB600-897-img0005.jpg "Western Blot: DNA-PKcs [p Thr2609] Antibody [NB600-897] - Lane 1: Fus1 untreated, Lane 2: Fus1 IR (20Gy, 4h), Lane 3: Fus1 DNAPK inhibitor + IR, Lane 4: MO59J (DNAPK-) untreated, Lane 5: MO59J IR, Lane 6: Fus1 untreated, Lane 7: Fus1 IR (20Gy, 4h), Lane 8: Fus1 DNAPK inhibitor + IR, Lane 9: MO59J untreated, Lane 10: MO59J IR. Lanes 1-5 are nuclear extract whereas lanes 6-10 are whole cell lysates. MO59J is a cell line that lacks DNA-PKcs. FUS1 is the matched cell line complemented with a chromosomal fragment containing the DNA-PKcs gene. Approximately 20 ug of lysate was run on SDS-PAGE and transferred onto nitrocellulose, followed by reaction with a 1:1,000 dilution of anti-DNAPKcs antibody. Detection occurred using a 1:5,000 dilution of HRP-labeled Goat anti-Rabbit IgG for 1 hour at room temperature. A chemiluminescence system was used for signal detection (Roche) using a 1 min exposure time.")
| Reactivity | HuSpecies Glossary |
| Applications | WB, ELISA, IP |
| Clonality | Polyclonal |
| Host | Rabbit |
| Conjugate | Unconjugated |
| Description | The product was affinity purified from monospecific antiserum by immunoaffinity purification. Antiserum was first purified against the phosphorylated form of the immunizing peptide. The resultant affinity purified antibody was then cross-adsorbed against the non-phosphorylated form of the immunizing peptide Store vial at -20C prior to opening. Aliquot contents and freeze at -20C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use. |
| Immunogen | DNA-PKcs [p Thr2609] antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to amino acids surrounding Thr 2609 of human DNA-PKcss. (Uniprot: P78527) |
| Modification | p Thr2609 |
| Localization | Nuclear |
| Specificity | DNA-PKcss pT2609 antibody is directed against the phosphorylated form of human DNA-PKcss at the pT2609 residue. The resultant affinity purified antibody was then cross-adsorbed against the non-phosphorylated form of the immunizing peptide. This phospho-specific polyclonal antibody reacts with phosphorylated pT2609 of human DNA-PKcss. Reactivity with non-phosphorylated human DNA-PKcss is minimal. A BLAST analysis was used to suggest reactivity with this protein from human and chimpanzee based on 100% homology for the immunogen sequence. However, cross-reactivity is expected with mouse, rat, dog, chicken and horse DNA-PKcss based on a high degree of homology to the immunogen sequence. Cross-reactivity with DNA-PKcss homologues from other sources has not been determined. |
| Isotype | IgG |
| Clonality | Polyclonal |
| Host | Rabbit |
| Gene | PRKDC |
| Purity | Immunogen affinity purified |
| Innovator's Reward | Test in a species/application not listed above to receive a full credit towards a future purchase. |
| Dilutions |
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| Application Notes | This product has been tested for use in ELISA, western blot, and IP. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 460 kDa in size corresponding to DNA PKcs by western blotting in the appropriate cell lysate or extract. Alternate splice variants have been described for this protein. Best western blotting results are seen when IP is performed prior to detection. This antibody detects an inducible signal at the correct height after DNA damage. However, the antibody may also detect an IR-inducible signal in cells lacking DNA PKcs (MO59J cells) at the same size as DNA PKcs if western blotting is performed directly. We believe that this additional band is 53BP1 that runs at the same size as DNA PKcs and is also phosphorylated in an IR-dependent manner. Similar results were seen with another phospho DNA PKcs antibody indicating that this result is general to antibodies to this phospho site, rather than specific to this antibody. |
| Storage | Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles. |
| Buffer | 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2 |
| Preservative | 0.01% Sodium Azide |
| Purity | Immunogen affinity purified |
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Secondary Antibodies |
Isotype Controls |
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