Anti-DNA PKcs抗体[18-2] (ab1832)

概述

  • 产品名称Anti-DNA PKcs抗体[18-2]
    参阅全部 DNA PKcs 一抗
  • 描述
    小鼠单克隆抗体[18-2] to DNA PKcs
  • 经测试应用适用于: Functional Studies, IHC-P, IHC-Fr, IP, WB, ICC/IF, Flow Cytmore details
  • 种属反应性
    与反应: Mouse, Rat, Human, Marmoset (common)
  • 免疫原

    Recombinant fragment corresponding to Human DNA PKcs.
    Database link: P78527

  • 表位Amino acids 1-2713.
  • 阳性对照
    • LS174T cells. Tonsil.

性能

应用

Our Abpromise guarantee covers the use of ab1832 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
Functional Studies Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.

Staining of formalin-fixed tissues requires boiling tissue sections in 10mM citrate buffer, pH 6.0,for 10-20 min followed by cooling at RT for 20 min.

IHC-Fr Use at an assay dependent concentration.
IP Use at an assay dependent concentration. (protein G) Use at a concentration of 2µg/mg of protein lysate (native and denatured).
WB Use at an assay dependent concentration. Detects a band of approximately 460 kDa (predicted molecular weight: 469 kDa).

This antibody reacts with intact DNA-PKcs as well as its degradation products.

ICC/IF Use at an assay dependent concentration. PubMed: 20709753
Flow Cyt Use at an assay dependent concentration.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

靶标

  • 功能Serine/threonine-protein kinase that acts as a molecular sensor for DNA damage. Involved in DNA nonhomologous end joining (NHEJ) required for double-strand break (DSB) repair and V(D)J recombination. Must be bound to DNA to express its catalytic properties. Promotes processing of hairpin DNA structures in V(D)J recombination by activation of the hairpin endonuclease artemis (DCLRE1C). The assembly of the DNA-PK complex at DNA ends is also required for the NHEJ ligation step. Required to protect and align broken ends of DNA. May also act as a scaffold protein to aid the localization of DNA repair proteins to the site of damage. Found at the ends of chromosomes, suggesting a further role in the maintenance of telomeric stability and the prevention of chromosomal end fusion. Also involved in modulation of transcription. Recognizes the substrate consensus sequence [ST]-Q. Phosphorylates 'Ser-139' of histone variant H2AX/H2AFX, thereby regulating DNA damage response mechanism. Phosphorylates DCLRE1C, c-Abl/ABL1, histone H1, HSPCA, c-jun/JUN, p53/TP53, PARP1, POU2F1, DHX9, SRF, XRCC1, XRCC1, XRCC4, XRCC5, XRCC6, WRN, MYC and RFA2. Can phosphorylate C1D not only in the presence of linear DNA but also in the presence of supercoiled DNA. Ability to phosphorylate p53/TP53 in the presence of supercoiled DNA is dependent on C1D.
  • 序列相似性Belongs to the PI3/PI4-kinase family.
    Contains 1 FAT domain.
    Contains 1 FATC domain.
    Contains 2 HEAT repeats.
    Contains 1 PI3K/PI4K domain.
    Contains 3 TPR repeats.
  • 翻译后修饰Phosphorylated upon DNA damage, probably by ATM or ATR. Autophosphorylated on Thr-2609, Thr-2638 and Thr-2647. Thr-2609 is a DNA damage-inducible phosphorylation site (inducible with ionizing radiation, IR). Autophosphorylation induces a conformational change that leads to remodeling of the DNA-PK complex, requisite for efficient end processing and DNA repair.
    S-nitrosylated by GAPDH.
  • 细胞定位Nucleus.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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
    • HYRC 1 antibody
    • HYRC antibody
    • HYRC1 antibody
    • IMD26 antibody
    • p350 antibody
    • p460 antibody
    • PKRDC antibody
    • PRKDC antibody
    • PRKDC_HUMAN antibody
    • Protein Kinase DNA Activated Catalytic Polypeptide antibody
    • XRCC 7 antibody
    • XRCC7 antibody
    see all

Anti-DNA PKcs antibody [18-2] 图像

  • ab1832 staining DNA PKcs in common marmoset small intestine tissue sections by Immunohistochemistry (Formalin/PFA-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 2% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/1000 in TBS/BSA/azide) for 2 hours at 21°C. A Biotin-conjugated goat anti-mouse IgG polyclonal (1/300) was used as the secondary antibody.

    See Abreview

  • Immunocytochemistry/Immunofluorescence analysis of mouse primary neurones labelling DNA PKcs with ab1832 at a dilution of 1/500. Cells were fixed with Formaldehyde and permeabilized with 0.5% Triton. The secondary antibody used was Alexa Fluor® 488 conjugated goat anti-mouse IgG at 1/1000.

    The image shows punctate DNA-pkc positivity in neuronal nuclei (green). The cytoplasm of the neurones is red (beta III tubulin detected by anti-Mouse. Nuclei were stained using Hoechst.

    This excellent image was kindly given to me by a Karolinska Institute BSc student (2016) Patrick Tannoury (doing his project at the Wolfson CARD Kings College London).

    See Abreview

  • Overlay histogram showing Jurkat cells stained with ab1832 (blue line). The cells were prepared as follows: Spin down, wash in FACS buffer 1x, fix, wash 2x, and stain with primary antibody overnight. Buffer was was 0.1% sodium azide with FBS in PBS. Cells fixed with paraformaldehyde and then permeabilized with Triton X-100 and NP40. Cell population was gated by isolating cell population from plot of SSC-A / FSA-A. The cells were then incubated the antibody at 1/100 for 24 hours. The secondary antibody used was FITC-conjugated Goat anti-Mouse polyclonal at 1/100 dilution. Isotype control antibody (red line) was used under the same conditions

    See Abreview

  • ab1832 staining DNA PKcs in human colon tissue sections by Immunohistochemistry (Formalin/PFA-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 2% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/1000 in TBS/BSA/azide) for 2 hours at 21°C. A Biotin-conjugated goat anti-mouse IgG polyclonal (1/300) was used as the secondary antibody.

    See Abreview

  • ab1832 staining DNA PKcs in HCT 116 cells by Immunocytochemistry/ Immunofluorescence.
    The cells were washed in PBS, fixed in 4% paraformaldehyde (in PBS) for 15 minutes, permeabilized in 0.3% Triton X-100 at room temperature for 10 minutes, followed by overnight incubation of ab1832 at a 1/500 dilution.
  • Anti-DNA PKcs antibody ab1832 used to stain Human tonsil tissue. The antibody requires antigen retrieval with citrate buffer and we recommend a 1:50-1:100 dilution with a 30 minute incubation time of the antibody at room temperature.

Anti-DNA PKcs antibody [18-2] (ab1832)参考文献

This product has been referenced in:
  • Keka IS  et al. Smarcal1 promotes double-strand-break repair by nonhomologous end-joining. Nucleic Acids Res N/A:N/A (2015). Human . Read more (PubMed: 26089390) »
  • Zhou Y  et al. Quantitation of DNA double-strand break resection intermediates in human cells. Nucleic Acids Res 42:e19 (2014). WB ; Human . Read more (PubMed: 24362840) »

See all 11 Publications for this product

Product Wall

Application Western blot
Sample Human Cell lysate - whole cell (U2OS cells)
Gel Running Conditions Reduced Denaturing (3-8% Gradient Gel)
Loading amount 50 µg
Specification U2OS cells
Blocking step Licor Blocking Buffer as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Oct 05 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (Primary neurones)
Permeabilization Yes - 0.5% Triton
Specification Primary neurones
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

提交于 Jul 13 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Rat Tissue sections (Small intestine)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Specification Small intestine
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

提交于 May 16 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Pig Tissue sections (Small intestine)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Specification Small intestine
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

提交于 May 16 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Lung)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Specification Lung
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

提交于 May 16 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Marmoset (common) Tissue sections (Small intestine)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Specification Small intestine
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

提交于 May 16 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Colon)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Specification Colon
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

提交于 May 16 2016

Application Immunohistochemistry (Frozen sections)
Blocking step 0.25%TritonX100, 0.2%Gelatine as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.2% · Temperature: 25°C
Sample Mouse Tissue sections (mouse embryonic spinal cord E11)
Specification mouse embryonic spinal cord E11
Permeabilization Yes - 0.25%TritonX100
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Aug 07 2014

DISCOUNT CODE: ***
Expiration date: November 6, 2012

This code will give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview forab1832 in ChIPand include this code in the “Additio...

Read More



- to optimize the transfer conditions by increasing the transfer time to 24 hours (in some examples, for big proteins, the transfer is done for 48 hours)
-a "boost" at 70-80V for 1 hour at the end of the transfer can be of sign...

Read More

1-10 of 14 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"