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76696
Non-Homologous End Joining (NHEJ) DNA Repair Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Non-Homologous End Joining (NHEJ) DNA Repair Antibody Sampler Kit #76696

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Non-Homologous End Joining (NHEJ) DNA Repair Antibody Sampler Kit: Image 1

Flow cytometric analysis of M059J cells (blue) and M059K cells (green) using DNA-PKcs (E6U3A) Rabbit mAb (solid lines) or a concentration matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Non-Homologous End Joining (NHEJ) DNA Repair Antibody Sampler Kit: Image 2

Western blot analysis of extracts from M059K (DNA-PKcs positive) and M059J (DNA-PKcs negative) cells, untreated (-), or treated with neocarzinostatin (0.5 µg/ml, 1 hr; +) to induce DNA double strand breaks, using Phospho-DNA-PKcs (Ser2056) (E9J4G) Rabbit mAb (upper), total DNA-PKcs (E6U3A) Rabbit mAb #38168 (middle), or α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).

Non-Homologous End Joining (NHEJ) DNA Repair Antibody Sampler Kit: Image 3

Western blot analysis of extracts from various cell lines using Ku70 (D10A7) Rabbit mAb.

Non-Homologous End Joining (NHEJ) DNA Repair Antibody Sampler Kit: Image 4

Confocal immunofluorescent analysis of HeLa cells using Ku80 (C48E7) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).

Non-Homologous End Joining (NHEJ) DNA Repair Antibody Sampler Kit: Image 5

Western blot analysis of extracts from various cell lines using DNA Ligase IV (D5N5N) Rabbit mAb.

Non-Homologous End Joining (NHEJ) DNA Repair Antibody Sampler Kit: Image 6

Western blot analysis of extracts of various cell lines using XLF Antibody.

Non-Homologous End Joining (NHEJ) DNA Repair Antibody Sampler Kit: Image 7

Immunoprecipitation of Artemis from HT-29 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Artemis (D7O8V) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot was performed using Artemis (D7O8V) Rabbit mAb.

Non-Homologous End Joining (NHEJ) DNA Repair Antibody Sampler Kit: Image 8

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

Non-Homologous End Joining (NHEJ) DNA Repair Antibody Sampler Kit: Image 9

Confocal immunofluorescent analysis of M059K cells (left, positive) and M059J cells (right, negative) using DNA-PKcs (E6U3A) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red).

Non-Homologous End Joining (NHEJ) DNA Repair Antibody Sampler Kit: Image 10

Immunohistochemical analysis of frozen SKOV-3 xenograft using Ku80 (C48E7) Rabbit mAb.

Non-Homologous End Joining (NHEJ) DNA Repair Antibody Sampler Kit: Image 11

Western blot analysis of extracts from various cell lines using Artemis (D7O8V) Rabbit mAb.

Non-Homologous End Joining (NHEJ) DNA Repair Antibody Sampler Kit: Image 12

Immunohistochemical analysis of paraffin-embedded human ductal breast carcinoma using DNA-PKcs (E6U3A) Rabbit mAb.

Non-Homologous End Joining (NHEJ) DNA Repair Antibody Sampler Kit: Image 13

Immunohistochemical analysis of paraffin-embedded human glioblastoma using Ku80 (C48E7) Rabbit mAb.

Non-Homologous End Joining (NHEJ) DNA Repair Antibody Sampler Kit: Image 14

Immunohistochemical analysis of paraffin-embedded normal human colon using DNA-PKcs (E6U3A) Rabbit mAb.

Non-Homologous End Joining (NHEJ) DNA Repair Antibody Sampler Kit: Image 15

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Ku80 (C48E7) Rabbit mAb.

Non-Homologous End Joining (NHEJ) DNA Repair Antibody Sampler Kit: Image 16

Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma using DNA-PKcs (E6U3A) Rabbit mAb.

Non-Homologous End Joining (NHEJ) DNA Repair Antibody Sampler Kit: Image 17

Immunohistochemical analysis of paraffin-embedded human melanoma using Ku80 (C48E7) Rabbit mAb.

Non-Homologous End Joining (NHEJ) DNA Repair Antibody Sampler Kit: Image 18

Immunohistochemical analysis of paraffin-embedded human lung squamous cell carcinoma using DNA-PKcs (E6U3A) Rabbit mAb.

Non-Homologous End Joining (NHEJ) DNA Repair Antibody Sampler Kit: Image 19

Immunohistochemical analysis of paraffin-embedded human GIST using Ku80 (C48E7) Rabbit mAb.

Non-Homologous End Joining (NHEJ) DNA Repair Antibody Sampler Kit: Image 20

Immunohistochemical analysis of paraffin-embedded human endometriod carcinoma using DNA-PKcs (E6U3A) Rabbit mAb.

Non-Homologous End Joining (NHEJ) DNA Repair Antibody Sampler Kit: Image 21

Western blot analysis of cell extracts from HeLa, A549 and COS cells using Ku80 (C48E7) Rabbit mAb.

Non-Homologous End Joining (NHEJ) DNA Repair Antibody Sampler Kit: Image 22

Immunohistochemical analysis of paraffin-embedded M059K cell pellet (left, positive) or M059J cell pellet (right, negative) using DNA-PKcs (E6U3A) Rabbit mAb.

Non-Homologous End Joining (NHEJ) DNA Repair Antibody Sampler Kit: Image 23

Western blot analysis of extracts from M059K cells, which express normal levels of DNA-PKcs, and M059J cells, which lack DNA-PKcs, using DNA-PKcs (E6U3A) Rabbit mAb (upper) or α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).

To Purchase # 76696T
Product # Size Price
76696T
1 Kit  (7 x 20 µl) N/A

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
DNA-PKcs (E6U3A) Rabbit mAb 38168 20 µl
  • WB
  • IHC
  • IF
  • F
H 450 Rabbit IgG
Phospho-DNA-PKcs (Ser2056) (E9J4G) Rabbit mAb 68716 20 µl
  • WB
H 450 Rabbit IgG
Ku70 (D10A7) Rabbit mAb 4588 20 µl
  • WB
H M R Mk 70 Rabbit IgG
Ku80 (C48E7) Rabbit mAb 2180 20 µl
  • WB
  • IP
  • IHC
  • IF
H Mk 86 Rabbit IgG
DNA Ligase IV (D5N5N) Rabbit mAb 14649 20 µl
  • WB
H 100 Rabbit IgG
XLF Antibody 2854 20 µl
  • WB
  • IP
H 39 Rabbit 
Artemis (D7O8V) Rabbit mAb 13381 20 µl
  • WB
  • IP
H Mk 90 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product Description

The Non-Homologous End Joining (NHEJ) DNA Repair Antibody Sampler Kit provides an economical means of detecting proteins involved in NHEJ DNA repair. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Each antibody in the Non-Homologous End Joining (NHEJ) DNA Repair Antibody Sampler Kit detects endogenous levels of its target protein. Phospho-DNA-PKcs (Ser2056) (E9J4G) Rabbit mAb recognizes endogenous levels of DNA-PKcs protein only when phosphorylated at Ser2056.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Pro608 of human DNA-PKcs protein, Val294 of mouse Ku70 protein, the carboxy terminus of human Ku80 protein, Leu771 of human DNA ligase IV protein, and Pro367 of human artemis protein. Phosphorylation-specific monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser2056 of human DNA-PKcs protein. Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to amino acids near the carboxy terminus of human XLF protein. Polyclonal antibodies are purified by peptide affinity chromatography.

Background

DNA double-strand breaks (DSBs) are potentially hazardous lesions that can be induced by ionizing radiation (IR), radiomimetic chemicals, or DNA replication inhibitors. Cells recognize and repair DSBs via two distinct but partly overlapping signaling pathways, non-homologous end joining (NHEJ) and homologous recombination (HR). DNA repair via the HR pathway is restricted to S and G2 phases of the cell cycle, while NHEJ can occur during any phase. NHEJ machinery is also utilized in V(D)J recombination, a process that generates diversity in immunoglobulin and T cell receptor genes. Defects in both pathways have been associated with human disease, including cancer (1).

DNA repair through the NHEJ pathway involves a core group of proteins that includes the Ku heterodimer (Ku70/Ku80), DNA-PKcs, DNA ligase IV, XRCC4, and XLF. XLF interacts with XRCC4 and promotes the ligation of DNA strands by DNA ligase IV and the ligase cofactor XRCC4. The ATP-dependent ligation of free DNA ends is the final step in the NHEJ repair pathway (2). DNA ligase IV and the endonuclease artemis suppress homologous recombination and promote NHEJ (3).

  1. Hartlerode, A.J. and Scully, R. (2009) Biochem J 423, 157-68.
  2. Tsai, C.J. et al. (2007) Proc Natl Acad Sci U S A 104, 7851-6.
  3. Kurosawa, A. et al. (2013) PLoS One 8, e72253.

Pathways & Proteins

Explore pathways + proteins related to this product.

Limited Uses

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For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
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