Skip to content

Phospho-DNA PKcs (S2056) Recombinant Rabbit Monoclonal Antibody [JE41-92]

Usd: 385

Specification

Bulk Order Quote | Customization Request

Catalog# HA722066

Phospho-DNA PKcs (S2056) Recombinant Rabbit Monoclonal Antibody [JE41-92]

Application

  • WB

  • IF-Cell

Reactivity

  • Human

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Phospho-DNA PKcs (S2056) Recombinant Rabbit Monoclonal Antibody [JE41-92]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide corresponding to residues surrounding Ser2056 of human DNA-PKcs protein.

Species Reactivity

Human

Validated Applications

WB, IF-Cell

Molecular Weight

Predicted band size: 469 kDa

Positive Control

HT-29 treated with UV for 3 hours cell lysate, HT-29 cells treated with UV for 3 hours.

Conjugation

unconjugated

Clone Number

JE41-92

RRID

AB_3096730

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IF-Cell

  • 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 consisting of 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. 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. DNA-PK also cooperates with ATR and ATM to phosphorylate proteins involved in the DNA damage checkpoint.

Background References

1. Dibitetto D et al. DNA-PKcs promotes fork reversal and chemoresistance. Mol Cell. 2022 Oct

2. Zou R et al. DNA-PKcs promotes sepsis-induced multiple organ failure by triggering mitochondrial dysfunction. J Adv Res. 2022 Nov

Subcellular Location

Nucleus, nucleolus.

UNIPROT

SwissProt: P78527 Human

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

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-DNA PKcs (S2056) on different lysates with Rabbit anti-Phospho-DNA PKcs (S2056) antibody (<a href="/products/HA722066" style="font-weight: bold;text-decoration: underline;">HA722066</a>) at 1/1,000 dilution.<br /><br />Lane 1: HT-29 cell lysate<br />Lane 2: HT-29 treated with UV for 3 hours cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 469 kDa<br />Observed band size: 469 kDa<br /><br />Exposure time: 2 minutes;<br /><br />6% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA722066" style="font-weight: bold;text-decoration: underline;">HA722066</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of Phospho-DNA PKcs (S2056) on different lysates with Rabbit anti-Phospho-DNA PKcs (S2056) antibody (HA722066) at 1/1,000 dilution.

    Lane 1: HT-29 cell lysate
    Lane 2: HT-29 treated with UV for 3 hours cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 469 kDa
    Observed band size: 469 kDa

    Exposure time: 2 minutes;

    6% 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 (HA722066) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Immunocytochemistry analysis of HT-29 cells treated with or without UV for 3 hours labeling Phospho-DNA PKcs (S2056) with Rabbit anti-Phospho-DNA PKcs (S2056) antibody (<a href="/products/HA722066" style="font-weight: bold;text-decoration: underline;">HA722066</a>) at 1/100 dilution.<br /><br />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 Rabbit anti-Phospho-DNA PKcs (S2056) antibody (<a href="/products/HA722066" style="font-weight: bold;text-decoration: underline;">HA722066</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    ☑ Cell treatment (CT)

    Immunocytochemistry analysis of HT-29 cells treated with or without UV for 3 hours labeling Phospho-DNA PKcs (S2056) with Rabbit anti-Phospho-DNA PKcs (S2056) antibody (HA722066) 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 Rabbit anti-Phospho-DNA PKcs (S2056) antibody (HA722066) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was 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"

Citation