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DNA-PKcs [p Thr2609] Antibody

Novus Biologicals, part of Bio-Techne | Catalog # NB600-897

Novus Biologicals, part of Bio-Techne
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NB600-897

Key Product Details

Validated by

Biological Validation

Species Reactivity

Human

Applications

ELISA, Immunoprecipitation, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Product Specifications

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)

Reactivity Notes

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.

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.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

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.

Scientific Data Images for DNA-PKcs [p Thr2609] Antibody

Western Blot: DNA-PKcs [p Thr2609] Antibody [NB600-897]

Western Blot: DNA-PKcs [p Thr2609] Antibody [NB600-897]

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.
DNA-PKcs [p Thr2609] Antibody

DNA-PKcs [p Thr2609] Antibody

Western Blot results of Rabbit DNA-PKcs [p Thr2609] Antibody. Lane 1: Molecular Weight Ladder. Lane 2: DNA Pkc phosphor control. Lane 3: DNA Pkc control. Lane 4: BSA. Load: 10ug. Primary Antibody: Rabbit Anti-DNA Pkcs pT609 at 1ug/mL overnight at 4C. Secondary Antibody: Goat anti-Rabbit HRP

Applications for DNA-PKcs [p Thr2609] Antibody

Application
Recommended Usage

ELISA

1:300000

Immunoprecipitation

1:10 - 1:500

Western Blot

1:500-1:2000
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.

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

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

Preservative

0.01% Sodium Azide

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles.

Background: DNA-PKcs

DNA PK is the key component of the non-homologous end-joining (NHEJ) pathway of DSB repair in mammalian cells. DNA PK consists of a heterodimeric DNA-binding subunit (Ku70/80) and an approximately 465 kDa catalytic subunit (DNA PKcs). DNA PKcs is a serine/threonine protein kinase whose activity is greatly stimulated by its recruitment to DNA breaks by the Ku heterodimer.

Long Name

DNA-dependent Protein Kinase Catalytic Subunit

Alternate Names

DNAPK, DNAPKcs, DNPK1, HYRC1, p350, PRKDC, XRCC7

Entrez Gene IDs

5591 (Human)

Gene Symbol

PRKDC

UniProt

Additional DNA-PKcs Products

Product Documents for DNA-PKcs [p Thr2609] Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for DNA-PKcs [p Thr2609] Antibody

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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