DNA-PKcs Recombinant antibody

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  WB analysis of HT-29 using 85370-2-RR

  HT-29 cells were subjected to SDS PAGE followed by western blot with 85370-2-RR (DNA-PKcs antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  WB analysis of HeLa using 85370-2-RR

  HeLa cells were subjected to SDS PAGE followed by western blot with 85370-2-RR (DNA-PKcs antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  IHC staining of human colon cancer using 85370-2-RR

  Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 85370-2-RR (DNA-PKcs antibody) at dilution of 1:1000 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human colon cancer using 85370-2-RR

  Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 85370-2-RR (DNA-PKcs antibody) at dilution of 1:1000 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of MCF-7 using 85370-2-RR

  Immunofluorescent analysis of (4% PFA) fixed MCF-7 cells using DNA-PKcs antibody (85370-2-RR, Clone: 242676E6 ) at dilution of 1:500 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-Phalloidin (red).

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  IF Staining of HepG2 using 85370-2-RR

  Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using DNA-PKcs antibody (85370-2-RR, Clone: 242676E6 ) at dilution of 1:500 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-Phalloidin (red).

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  IF Staining of HeLa using 85370-2-RR

  Immunofluorescent analysis of (4% PFA) fixed HeLa cells using DNA-PKcs antibody (85370-2-RR, Clone: 242676E6 ) at dilution of 1:500 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-Phalloidin (red).

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  FC experiment of HepG2 using 85370-2-RR

  1x10^6 HepG2 cells were intracellularly stained with 0.25 ug DNA-PKcs Recombinant antibody (85370-2-RR, Clone:242676E6) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2)(red), or 0.25 ug Rabbit IgG Isotype Control RecAb (98136-1-RR, Clone: 240953C9) (blue). Cells were fixed and permeabilized with Transcription Factor Staining Buffer Kit (PF00011).

DNA-PKcs Uni-rAb^TM Recombinant Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Cat No. 85370-2-RR

Clone No.242676E6

Host / Isotype

Rabbit / IgG

Reactivity

human

Applications

WB, IHC, IF/ICC, FC (Intra), ELISA

Synonyms

DNAPK, DNPK1, PRKDC, DNA-PK catalytic subunit, DNA-dependent protein kinase catalytic subunit

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Tested Applications

• Positive WB detected in: HeLa cells, HT-29 cells
• Positive IHC detected in: human colon cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: HeLa cells, MCF-7 cells, HepG2 cells
• Positive FC (Intra) detected in: HepG2 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:500-1:2000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:250-1:1000
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.25 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

85370-2-RR targets DNA-PKcs in WB, IHC, IF/ICC, FC (Intra), ELISA applications and shows reactivity with human samples.

• Tested Reactivity: human
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: Peptide
• Full Name: protein kinase, DNA-activated, catalytic polypeptide
• Calculated Molecular Weight: 469 kDa
• Observed Molecular Weight: 350-460 kDa
• GenBank Accession Number: NM_006904
• Gene Symbol: PRKDC
• Gene ID (NCBI): 5591
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• UNIPROT ID: P78527
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

PRKDC, also named as HYRC, HYRC1, DNPK1 and p460, belongs to the PI3/PI4-kinase family. PRKDC is a serine/threonine-protein kinase that acts as a molecular sensor for DNA damage. Involved in DNA nonhomologous end joining (NHEJ), PRKDC is required for double-strand break (DSB) repair and V(D)J recombination. PRKDC must be bound to DNA to express its catalytic properties. It promotes processing of hairpin DNA structures in V(D)J recombination by activation of the hairpin endonuclease artemis (DCLRE1C). It is required to protect and align broken ends of DNA. PRKDC may also act as a scaffold protein to aid the localization of DNA repair proteins to the site of damage. It is found at the ends of chromosomes, suggesting a further role in the maintenance of telomeric stability and the prevention of chromosomal end fusion. It also involved in modulation of transcription. It recognizes the substrate consensus sequence [ST]-Q. PRKDC phosphorylates 'Ser-139' of histone variant H2AX/H2AFX, thereby regulating DNA damage response mechanism.