Anti-DNA PKcs (phospho T2609) antibody (ab4194)
- Product nameAnti-DNA PKcs (phospho T2609) antibodySee all DNA PKcs primary antibodies ...
- DescriptionRabbit polyclonal to DNA PKcs (phospho T2609)
- SpecificityThis phospho specific polyclonal antibody reacts with phosphorylated pT2609 of human DNA PKcs. Reactivity with non-phosphorylated human DNA PKcs is minimal by ELISA. This antibody recognises phospho T2609 DNA PKcs as demonstrated: IPed DNA PKcs (using another DNA PKcs antibody) blotted with the pT2609 antibody shows an inducible signal at the correct height after DNA damage. However, unfortunately the antibody picks up an IR-inducible signal in cells lacking DNA-PKcs (MO59J cells) at the same size as DNA-PKcs. We believe that this is 53BP1 which 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. This antibody may be a useful marker for DNA damage. It may also still be useful in the study of phospho DNA PKcs T2609.
- Tested applicationsICC/IF, WB more details
- Species reactivityReacts with: Human
Synthetic peptide: PMFVEpTQASQG, corresponding to amino acids 2604-2614 of Human DNA PKcs (phosphorylated form of human DNA PKcs at the pT2609 residue).
- Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferPreservative: 0.01% Sodium Azide
Constituents: 0.15M Sodium Chloride, 0.02M Potassium Phosphate. pH 7.2
- Concentration information loading...
- PurityImmunogen affinity purified
- Purification notesThe 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. The product is sterile filtered.
- Clonality Polyclonal
- Research Areas
Our Abpromise guarantee covers the use of ab4194 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||ICC/IF: Use at an assay dependent dilution.|
|WB||WB: 1/1000. Detects a band of approximately >400 kDa (predicted molecular weight: 469 kDa). Works in WB after IP with another DNA PKcs antibody. When used without IP, this antibody also detects another phospho protein, believed to be 53BP1.Caution: when used on whole cell extracts (without IP) this antibody detects several background bands that are not seen with nuclear extracts.|
- FunctionSerine/threonine-protein kinase that acts as a molecular sensor for DNA damage. Involved in DNA nonhomologous end joining (NHEJ) required for double-strand break (DSB) repair and V(D)J recombination. Must be bound to DNA to express its catalytic properties. Promotes processing of hairpin DNA structures in V(D)J recombination by activation of the hairpin endonuclease artemis (DCLRE1C). The assembly of the DNA-PK complex at DNA ends is also required for the NHEJ ligation step. Required to protect and align broken ends of DNA. May also act as a scaffold protein to aid the localization of DNA repair proteins to the site of damage. Found at the ends of chromosomes, suggesting a further role in the maintenance of telomeric stability and the prevention of chromosomal end fusion. Also involved in modulation of transcription. Recognizes the substrate consensus sequence [ST]-Q. Phosphorylates 'Ser-139' of histone variant H2AX/H2AFX, thereby regulating DNA damage response mechanism. Phosphorylates DCLRE1C, c-Abl/ABL1, histone H1, HSPCA, c-jun/JUN, p53/TP53, PARP1, POU2F1, DHX9, SRF, XRCC1, XRCC1, XRCC4, XRCC5, XRCC6, WRN, MYC and RFA2. Can phosphorylate C1D not only in the presence of linear DNA but also in the presence of supercoiled DNA. Ability to phosphorylate p53/TP53 in the presence of supercoiled DNA is dependent on C1D.
- Sequence similaritiesBelongs to the PI3/PI4-kinase family.
Contains 1 FAT domain.
Contains 1 FATC domain.
Contains 2 HEAT repeats.
Contains 1 PI3K/PI4K domain.
Contains 3 TPR repeats.
modificationsPhosphorylated upon DNA damage, probably by ATM or ATR. Autophosphorylated on Thr-2609, Thr-2638 and Thr-2647. Thr-2609 is a DNA damage-inducible phosphorylation site (inducible with ionizing radiation, IR). Autophosphorylation induces a conformational change that leads to remodeling of the DNA-PK complex, requisite for efficient end processing and DNA repair.
S-nitrosylated by GAPDH.
- Cellular localizationNucleus.
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Anti-DNA PKcs (phospho T2609) antibody images
Predicted band size : 469 kDa
Western blot on cell lysates using ab4194 at 1/1000.
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
Lane 1-5: Nuclear Lysate
Lane 6-10: Whole Cell Lysate
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.
As discussed above, we believe the upper bands represent a mixture of DNA PKcs phosphorylation and 53BP1 phosphorylation.
References for Anti-DNA PKcs (phospho T2609) antibody (ab4194)
This product has been referenced in:
- Bencokova Z et al. ATM activation and signaling under hypoxic conditions. Mol Cell Biol 29:526-37 (2009). WB ; Human . Read more (PubMed: 18981219) »
- Roig AI et al. Two- and three-dimensional models for risk assessment of radiation-enhanced colorectal tumorigenesis. Radiat Res 171:33-40 (2009). ICC/IF ; Human . Read more (PubMed: 19138051) »