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Primary Antibodies > Phospho Specific Antibodies

DNA PKcs pT 2609 Antibody

Rabbit Polyclonal

Code

600-401-494

Size

100 µg

Price

$294.00

Availability

Ships next business day.

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Western blot using Rockland's Affinity Purified anti-DNAPKcs antibody shows detection of a 460 kDa band corresponding to human DNAPKcs in various preparations. 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 µg 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.

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Synonyms: DNA dependent protein kinase catalytic subunit antibody, DNA PK catalytic subunit antibody, DNAPK antibody, DNAPK catalytic subunit antibody, DNPK 1 antibody, DNPK1 antibody, HYRC 1 antibody

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DNA PKcs pT 2609 Antibody Properties

Description

Anti-DNA PKcs pT 2609 (RABBIT) Antibody - 600-401-494

Host

Rabbit

Target Species

Human

Known Cross Reactivity

human, chimpanzee

Clonality

Polyclonal

Application

ELISA : 1:300,000
Western Blot : 1:500 - 1:2,000
Other Dilution: User Optimized

Physical State

Liquid (sterile filtered)

Shipping condition

Dry Ice

Format

IgG

Label

Unconjugated

Concentration

0.93 mg/mL by UV absorbance at 280 nm

Buffer

0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2

Preservative

0.01% (w/v) Sodium Azide

Stabilizer

None

GeneName

PRKDC

DNA PKcs pT 2609 Antibody Description

Background

DNA dependent Protein Kinase (also called DNAPK, DNPK1, HYRC1, Protein Kinase DNA Activated Catalytic Polypeptide, XRCC7 and p460) consists of the 460 kDa DNA PKcs and a heterodimeric regulatory complex comprised of p70 Ku and p80 Ku (Ku autoantigen). DNA PKcs is a nuclear protein serine/threonine kinase present in a wide variety of eukaryotic species. DNA PKcs phosphorylates transcription factors, Sp1, Oct-1, p53 and SV40 large T antigen. DNA PKcs is involved in repairing double stranded DNA breaks. At the onset of apoptosis, DNA PKcs is rapidly inactivated by cleavage of the catalytic subunit into smaller polypeptides. Proteolysis of DNA PKcs is inhibited by the cysteine protease inhibitors iodoacetamide and N-ethylmaleimide. Alternative splicing can occur for this protein to produce at least two isoforms.

Immunogen

This affinity purified 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 PKcs.

Immunogen Type

Peptide

Post Translational Modification

Phosphorylation

Storage Condition

Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C 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 4° C as an undiluted liquid. Dilute only prior to immediate use.

Application Note

This affinity purified antibody has been tested for use in ELISA and by western blot. 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.

Purity/Specifity

This affinity purified antibody is directed against the phosphorylated form of human DNA PKcs at the pT2609 residue. 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. This phospho-specific polyclonal antibody reacts with phosphorylated pT2609 of human DNA PKcs. Reactivity with non-phosphorylated human DNA PKcs 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 PKcs based on a high degree of homology to the immunogen sequence. Cross-reactivity with DNA PKcs homologues from other sources has not been determined.

Disclaimer Note-General

This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 326, Gilbertsville, Pennsylvania, USA.

General Reference

Sawchuk,D.J., et al. (2004) Ku70/Ku80 and DNA-dependent protein kinase catalytic subunit modulate RAG-mediated cleavage: implications for the enforcement of the 12/23 rule. J. Biol. Chem. 279 (28), 29821-29831. Graham,K.L., Gustin,K.E., Rivera,C., Kuyumcu-Martinez,N.M., Choe,S.S., Lloyd,R.E., Sarnow,P. and Utz,P.J. (2004) Proteolytic cleavage of the catalytic subunit of DNA-dependent protein kinase during poliovirus infection. J. Virol. 78 (12), 6313-6321. Panta,G.R., Kaur,S., Cavin,L.G., Cortes,M.L., Mercurio,F., Lothstein,L., Sweatman,T.W., Israel,M. and Arsura,M. (2004) ATM and the catalytic subunit of DNA-dependent protein kinase activate NF-kappaB through a common MEK/extracellular signal-regulated kinase/p90(rsk) signaling pathway in response to distinct forms of DNA damage. Mol. Cell. Biol. 24 (5), 1823-1835.

Specific Reference

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