Anti-KAP1 antibody

• WB

Supplier Data

Western blot - Anti-KAP1 antibody (AB10484)

Cells prepared using NETN lysis buffer. Chemiluminescence detection.

All lanes:

Western blot - Anti-KAP1 antibody (ab10484) at 0.1 µg/mL

Lane 1:

HeLa whole cell lysate at 50 µg

Lane 2:

293T whole cell lysate at 50 µg

Lane 3:

3T3 whole cell lysate at 50 µg

Predicted band size: 88 kDa

false

Exposure time: 1s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KAP1 antibody (AB10484)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian carcinoma labeling KAP1 with ab10484 at 1/5000 dilution (0.2 μg/ml). DAB detection, Hematoxylin counterstain.

• IP

Supplier Data

Immunoprecipitation - Anti-KAP1 antibody (AB10484)

CTCF was immunoprecipitated from 1 mg HeLa whole cell lysate with ab10484 at 6 μg per reaction. Western blot was performed on the immunoprecipitate using ab10484 at 1 μg/mL.

Lysates prepared using NETN lysis buffer.

Lane 1 : ab10484 IP in HeLa whole cell lysate.

Lane 2 : Contol IgG in HeLa whole cell lysate.

Detection : Chemiluminescence.

Exposure time : 3 seconds.

All lanes:

Immunoprecipitation - Anti-KAP1 antibody (ab10484)

Predicted band size: 88 kDa

false

• IHC-P

AbReview24025****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KAP1 antibody (AB10484)

ab10484 staining KAP1 in murine coronal brain tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with paraformaldehyde and permeabilized using 1% Triton. Samples were then blocked with 5% serum for 1 hour at 25°C followed by incubation with the primary antibody at 2μg/ml for 16 hours at 4°C. An Alexa-Fluor 488-conjugated donkey anti-rabbit polyclonal was used as secondary antibody at a 1/1000 dilution.

Dentate gyrus of the hippocampus immunostained with DAPI (top) and KAP1 (bottom), which nicely labels the nuclei of the entire dentate gyrus.

Image courtesy of an anonymous Abreview.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KAP1 antibody (AB10484)

Immunohistochemical analysis of formalin-fixed, paraffin-embedded mouse plasmacytoma, labeling KAP1 with ab10484 at 1 μg/mL. Detection : DAB.

• WB

CiteAb

Western blot - Anti-KAP1 antibody (AB10484)

Western Blotting using Anti-KAP1 antibody, ab10484. Publication image from Liu, X. et al., 2017, Nat Commun, 28051062. Legend direct from paper.

PAXX and XLF have distinct functions during NHEJ.(a) Southern blot analyses of the product and intermediates of pMX-INV V(D)J recombination reporter in WT, Xlf−/− and Paxxδ/δ cells with or without ATM kinase inhibitor (KU55933, 15 µM) or DNA-PKcs inhibitor (NU7441, 5 µM) added 1 h before STI571 (3 µM). (b) Western blot for phosphorylated KAP1, total KAP1 and β-actin in total lysate from pre-B cells immediately after irradiation (at 1, 5 or 10 Gy). (c) Extrachromosomal SJ formation efficiencies in WT, Paxxδ/δ, Xlf−/− and Xlf−/−Paxxδ/δ Pre-B cells. Each box represents the average and s.d. of four independent repeats. Box centre line is plotted at the median and the box extends from the 25th to 75th percentiles. Whiskers go down to the smallest value and up to the largest value. The raw data were summarized in Supplementary Fig. 5A. The P value was calculated by two-tailed Student's t-test. (d) Laser-induced recruitment of GFP-PAXX or DsRed-PCNA in immortalized Ku80−/− MEFs with or without ectopic expression of full-length (FL) Ku80 or C-terminal truncated Ku80 (δCTD). Ten cells were tested for each genotype and representive images are shown here. (e) Laser-induced recruitment of GFP-Ku70 in immortalized WT, Xlf−/−, Paxx−/− and Xlf−/−Paxx−/− MEFs 1 min after damage. P<0.001 between Paxx−/− and WT cells, P<0.05 between Xlf−/−Paxx−/− and WT cells, two-way ANOVA (f) Laser-induced recruitment of GFP-Lig4 in immortalized WT, Xlf−/−, Paxx−/− and Xlf−/−Paxx−/− MEFs 1 min after damage. P<0.001 between Xlf−/− and WT cells, P<0.01 between Paxx−/− and WT cells, P<0.001 between Xlf−/−Paxx−/− and WT cells, two-way ANOVA. The bar graphs of e,f represent the average and error bars stand for s.d. (see Methods for details), same experiments were repeated for at least two times. The scale bar in d stands for 10 µm. (g) Working model. In this model, we propose that XLF (diagrammed as red bean sprout shape) enhances the efficient accumulation of Lig4 in complex with XRCC4 at the DNA ends, while PAXX (diagrammed as green bean sprout shape) promotes the accumulation of KU at the DNA ends. Together PAXX and XLF promote efficient DNA end-ligation through distinct mechanisms. XR4, XRCC4; Art, Artemis.

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