Anti-53BP1 antibody

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-53BP1 antibody (AB172580)

Immunocytochemistry/Immunofluorescence analysis of U2OS cells labelling 53BP1 with ab172580 at 1 : 1000. Cells were fixed with PFA and permeabilized with methanol for 2 minutes. Cells were incubated with the primary anitbody overnight at room temperature.

• Flow Cyt

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Flow Cytometry - Anti-53BP1 antibody (AB172580)

Flow Cytometry analysis of HeLa cells labelling 53BP1 with ab172580 at 1 : 800. Cells were fixed with 1% PFA. Cells were incuabted with the primary antibody overnight at 4°C.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-53BP1 antibody (AB172580)

Immunohistochemistry (formalin/PFA-fixed paraffin-embedded) analysis of Human linfoid tissue labelling 53BP1 with ab172580 at 1 : 1000. Antigen retrieval was by microwave boiling for 7 minutes in 10mM TrisHCl, pH9.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-53BP1 antibody (AB172580)

Immunocytochemistry/Immunofluorescence analysis of U2OS cells labelling 53BP1 with ab172580 at 1 : 1000. Cells were fixed with PFA and permeabilized with methanol for 2 minutes. Cells were incubated with the primary anitbody overnight at room temperature. Top - 53BP1, Middle - DAPI, Bottom - Merge.

• IP

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Immunoprecipitation - Anti-53BP1 antibody (AB172580)

Immunoprecipitation analysis of U2OS cells labelling 53BP1 with ab172580. Protein A beads were crosslinked to the serum.

All lanes:

Immunoprecipitation - Anti-53BP1 antibody (ab172580)

Predicted band size: 214 kDa

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• WB

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Western blot - Anti-53BP1 antibody (AB172580)

All lanes:

Western blot - Anti-53BP1 antibody (ab172580)

Lane 1:

U2OS (Control si)

Lane 2:

U2OS (53BP1 si)

Lane 3:

NIH 3T3 cell lysate at 40 µg

Lane 4:

PC12 cell lysate at 40 µg

Predicted band size: 214 kDa

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• ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-53BP1 antibody (AB172580)

Immunocytochemistry-immunofluorescence using Anti-53BP1 antibody, ab172580. Publication image from Ferreira-Gonzalez, S. et al., 2018, Nat Commun, 29523787. Legend direct from paper.

Paracrine senescence in the model is TGFβ-dependent. Mouse data is presented at day 2 after final tamoxifen administration (N = 6 per group). (Left) K19Cre-positive mice without tamoxifen injection. (Center) K19Cre-negative mice after tamoxifen injection. (Right) K19Cre-positive mice after tamoxifen injection. a p53 (red, indicative of primary senescence events in the cholangiocytes) and p21 (green) co-localize in the bile ducts. However, p21-positive p53-negative cholangiocytes were observed. b p21 (green) co-localizes with HNF4α-positive hepatocytes (red). c p27 (green) colocalizes with p53-positive cholangiocytes (red). p27 is expressed in cholangiocytes and hepatocytes in a paracrine manner. d p27 (green) co-localizes with HNF4α-positive hepatocytes (red). e Markers of senescence (such as 53BP1, H2A.X and DCR2, in red) are expressed in cholangiocytes and liver parenchyma, while expression of p53 (green) is restricted to the cholangiocytes. Single channels for this figure are provided in Supplementary Fig. 8a. Scale bars = 50 µm. f qRT-PCR analysis of Tgfb1 in the isolated bile ducts of Cre− +TM (N = 5) and induced Cre+ +TM (N = 6) K19-Mdm2flox/floxtdTomLSL mice day 2 after induction. *** denotes p < 0.005 (Mean ± SEM). Mann−Whitney test. g Analysis of common SASP’s factors by qRT-PCR (from left to right, Tgfbr1, Tgfbr2, Nfkb, Il1a and Il6). * denotes p < 0.05, ** p < 0.01 (Mean ± SEM). Mann−Whitney test. h Western blot of phospho-NF-κB-p65 and pSmad2/3. Each band represents the bile ducts isolated from one mouse. Blots are companied by at least one marker position (molecular weights MW, in kDa). Far right, western blot densitometry quantification (normalized to bActin control) show increased expression of NF-κBp-65 and pSmad2/3 after induction of the model. * denotes p < 0.05 (Mean ± SEM). Mann−Whitney test. i qRT-PCR of Nfkb in hepatocytes isolated from Cre− and Cre+ K19-Mdm2flox/floxtdTomLSL mice at day 2 after last tamoxifen administration (N = 5-6 per group). ** denotes p < 0.01 (Mean ± SEM). Mann−Whitney test

• WB

CiteAb

Western blot - Anti-53BP1 antibody (AB172580)

53BP1 western blot using anti-53BP1 antibody ab172580. Publication image and figure legend from Warmerdam, D. O., Alonso-de Vega, I., et al., 2020, EMBO Rep, PubMed 31782600.

ab172580 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab172580 please see the product overview.

PHF6 controls classical NHEJA, BU2OS cells expressing GFP-PHF6 and mCherry-NBS1 were locally irradiated in the absence and presence of ATM and ATR inhibitors. Shown is the recruitment of GFP-PHF6 (A) or mCherry-NBS1 (B) to laser-induced breaks in time. At least 30 cells were analyzed in three individual experiments, and error bars represent the SD.CU2OS-DR cells containing the I-SceI HR reporter were transfected with the indicated siRNA oligos and I-SceI. 48 h later, the cells were fixed and analyzed by flow cytometry. Represented is the relative repair efficiency as compared to the Luciferase control. Error bars represent the SEM of three independent experiments. Statistical significance was determined using a two-tailed, unpaired t-test (**p < 0.01, ***p < 0.001, ****p < 0.0001).DU2OS cells depleted of PHF6 with three different siRNA oligonucleotides and U2OS WT and PHF6 knockout cells were lysed and analyzed using Western blot with the indicated antibodies.EU2OS WT and PHF6 knockout cells transfected with siRNA oligos against luciferase or XRCC4 were lysed and analyzed using Western blot with the indicated antibodies. The remaining PHF6 signal in the PHF6 KO sample is likely due to a cross reaction at similar height as PHF6 (also see Fig EV3C).F, GDistribution of the deletion sizes (F) or the extent of microhomology for the category simple deletions (G) obtained from independently obtained I-SceI repair events (described in Fig 4E) in the indicated samples.

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