DNA-PKcs/PRKDC Recombinant Mouse Monoclonal Antibody [2-B3-R]
Catalog# HA601232
DNA-PKcs/PRKDC Recombinant Mouse Monoclonal Antibody [2-B3-R]
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WB
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IHC-P
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IF-Cell
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Human
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unconjugated
Overview
Product Name
DNA-PKcs/PRKDC Recombinant Mouse Monoclonal Antibody [2-B3-R]
Antibody Type
Recombinant Mouse Monoclonal Antibody
Immunogen
Recombinant protein within Mouse DNA-PKcs/PRKDC aa 3,900-4,128/4,128.
Species Reactivity
Human
Validated Applications
WB, IHC-P, IF-Cell
Target Molecular Weight
Predicted band size: 469 kDa
Positive Control
K-562 cell lysate, HepG2 cell lysate, HeLa cell lysate, Jurkat cell lysate, 293T cell lysate, HT-29 cell lysate, A549 cell lysate, MCF7 cell lysate, human colon cancer tissue, HepG2.
Conjugation
unconjugated
Clone Number
2-B3-R
RRID
Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG1
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:2,000
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IHC-P
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1:500
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IF-Cell
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1:100
Target
Function
DNA-dependent protein kinase, catalytic subunit, also known as DNA-PKcs, is an enzyme that in humans is encoded by the gene designated as PRKDC or XRCC7. DNA-PKcs belongs to the phosphatidylinositol 3-kinase-related kinase protein family. The DNA-Pkcs protein is a serine/threonine protein kinase comprising a single polypeptide chain of 4,128 amino acids. DNA-PKcs is the catalytic subunit of a nuclear DNA-dependent serine/threonine protein kinase called DNA-PK. The second component is the autoimmune antigen Ku. On its own, DNA-PKcs is inactive and relies on Ku to direct it to DNA ends and trigger its kinase activity. DNA-PKcs is required for the non-homologous end joining (NHEJ) pathway of DNA repair, which rejoins double-strand breaks. It is also required for V(D)J recombination, a process that utilizes NHEJ to promote immune system diversity. DNA-PKcs knockout mice have severe combined immunodeficiency due to their V(D)J recombination defect. Many proteins have been identified as substrates for the kinase activity of DNA-PK. Autophosphorylation of DNA-PKcs appears to play a key role in NHEJ and is thought to induce a conformational change that allows end processing enzymes to access the ends of the double-strand break.[9] DNA-PK also cooperates with ATR and ATM to phosphorylate proteins involved in the DNA damage checkpoint.
Background References
1. "Frameshift mutation in PRKDC, the gene for DNA-PKcs, in the DNA repair-defective, human, glioma-derived cell line M059J."Anderson C.W., Dunn J.J., Freimuth P.I., Galloway A.M., Allalunis-Turner M.J.Radiat. Res. 156:2-9(2001)
2. "Human DNA-activated protein kinase (DNA-PK) is homologous to phosphatidylinositol kinases."Poltoratsky V.P., Shi X., York J.D., Lieber M.R., Carter T.H.J. Immunol. 155:4529-4533(1995)
Subcellular Location
Nucleus, nucleolus.
UNIPROT
Synonyms
DNA dependent protein kinase catalytic subunit antibody
DNA PK catalytic subunit antibody
DNA-dependent protein kinase catalytic subunit antibody
DNA-PK catalytic subunit antibody
DNA-PKcs antibody
DNAPK antibody
DNAPK catalytic subunit antibody
DNPK 1 antibody
DNPK1 antibody
Hyper radiosensitivity of murine scid mutation, complementing 1 antibody
ExpandImages
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Western blot analysis of DNA-PKcs/PRKDC on different lysates with Mouse anti-DNA-PKcs/PRKDC antibody (HA601232) at 1/2,000 dilution.
Lane 1: K-562 cell lysate
Lane 2: HepG2 cell lysate
Lane 3: HeLa cell lysate
Lane 4: Jurkat cell lysate
Lane 5: 293T cell lysate
Lane 6: HT-29 cell lysate
Lane 7: A549 cell lysate
Lane 8: MCF7 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 469 kDa
Observed band size: 469 kDa
Exposure time: 23 seconds;
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 (HA601232) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Mouse anti-DNA-PKcs/PRKDC antibody (HA601232) at 1/500 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 (HA601232) at 1/500 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. -
Immunocytochemistry analysis of HepG2 cells labeling DNA-PKcs/PRKDC with Mouse anti-DNA-PKcs/PRKDC antibody (HA601232) 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 Mouse anti-DNA-PKcs/PRKDC antibody (HA601232) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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 (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"