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53bp1 nb100 304

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53BP1 (NB100-304) is an antibody product offered by Novus Biologicals. It is designed for use in research applications. The core function of this product is to recognize and bind to the 53BP1 protein, which is involved in the cellular response to DNA double-strand breaks.

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Lab products found in correlation

Lsm 780 confocal microscope, by Zeiss (3 mentions) γh2ax, by Merck Group (2 mentions) Brca1 d9, by Santa Cruz Biotechnology (1 mentions) Uhrf1, by BD (1 mentions) Sc 8017, by Santa Cruz Biotechnology (1 mentions) Axiocam 702, by Zeiss (1 mentions) Oct 3 4 sc 5279, by Santa Cruz Biotechnology (1 mentions) Axio imager m2, by Zeiss (1 mentions) Prolong gold antifade reagent, by Thermo Fisher Scientific (1 mentions) Zscan4c, by Merck Group (1 mentions) H3k36me2, by Active Motif (1 mentions) H4k16ac, by Active Motif (1 mentions) Xpb s 19, by Santa Cruz Biotechnology (1 mentions) H4k20me2, by Active Motif (1 mentions) Ha c29f4, by Cell Signaling Technology (1 mentions) Ring1b clone d22f2, by Cell Signaling Technology (1 mentions) Flag tag (flag), by Merck Group (1 mentions) Alexa fluor 568, by Thermo Fisher Scientific (1 mentions) γh2ax 05 636, by Merck Group (1 mentions) Alexa fluor 488, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

NB100-304 (71 citations) Anti-53BP1 (NB100-304, (6 citations) 53BP1 (NB100-304) and FANCD2 (NB100-182) (2 citations) Rabbit anti-53BP1 antibody (clone NB100-304, (2 citations)

5 protocols using 53bp1 nb100 304

1

Antibody-based Protein Characterization Protocol

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The following antibodies were used in this study: UHRF1 (#612264, dilution: 1:1,000, BD Science) for western and IP and (H-8, dilution: 1:1000, Santa Cruz) for IF. BRCA1 (D-9, dilution: 1:100, Santa Cruz) for IF and (C-20, 1:200, Santa Cruz) for western blot and IP. RIF1 (A300-569A, dilution: 1:1,000, Bethyl Laborataries), HA (H9658, dilution: 1:1,000, Sigma), FLAG (F3165, dilution: 1:1,000, sigma), ubiquitin (SC-8017, dilution: 1:500, Santa Cruz), γ-H2AX (05-636, dilution: 1:500, Millipore), RPA (ab2175, dilution: 1:200, Abcam), RAD51(GTX70230, dilution: 1:200, GeneTex), 53BP1(NB100-304, dilution: 1:1,000, Novus), Phospho-(Ser) CDKs substrate antibody (9477s, dilution: 1:500, CST), cyclin A antibody (SC751, dilution: 1:200, Santa Cruz). CtIP (61141, dilution: 1:500, Active Motif). UHRF1, BRCA1 and RIF1 cDNAs were subcloned into the Flag-tagged vector (pIRES2-EGFP) HA-tagged vector (pCDNA3.1-HA or pCMV-HA). All mutants were generated by site-directed mutagenesis and confirmed by sequencing.
Zhang H., Liu H., Chen Y., Yang X., Wang P., Liu T., Deng M., Qin B., Correia C., Lee S., Kim J., Sparks M., Nair A.A., Evans D.L., Kalari K.R., Zhang P., Wang L., You Z., Kaufmann S.H., Lou Z, & Pei H. (2016). A cell cycle-dependent BRCA1–UHRF1 cascade regulates DNA double-strand break repair pathway choice. Nature Communications, 7, 10201.
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2

Immunofluorescence and IF-FISH Protocols

Cited 1 time
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Immunofluorescence and IF–FISH were performed as previously described6 (link). In brief, cells fixed with 2% paraformaldehyde were incubated with the following primary antibodies, all at 1:1,000 dilution: OCT3/4 (sc-5279, Santa Cruz Biotechnology), p-KAP1 (A300-767A, Bethyl) or ZSCAN4C (AB4340, Millipore Sigma), γH2AX (05–636, Millipore), 53BP1 (NB100-304, Novus Biological). For IF–FISH, following secondary antibody staining, cells were fixed, denatured at 72 °C and hybridized with the AlexaFluor 488-TelC (TAACCC) PNA probe (PNA Bio). Slides were then mounted using ProLong Gold antifade reagent (Life Technologies), and images were acquired using a Zeiss Axio Imager M2 and an Axiocam 702 camera and ZEN 2.6 (blue edition) software. For ES cells, Z-stack images were acquired and displayed as maximum intensity projections. Final figures were assembled using Adobe Photoshop CC 2020 versions 21.0.1.47 and 21.2.1.265 and Adobe Illustrator 2020 24.2. Complete details regarding the number of cells analysed for each figure are provided in Supplementary Table 2.
Markiewicz-Potoczny M., Lobanova A., Loeb A.M., Kirak O., Olbrich T., Ruiz S, & Denchi E.L. (2020). TRF2-mediated telomere protection is dispensable in pluripotent stem cells. Nature, 589(7840), 110-115.
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3

Comprehensive Protein Marker Profiling

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The following antibodies were used: keratin 14AB7800, keratin 18 AB668, laminin AB11575, ATM AB78, pChk2 AB85743, TRF2 AB13579, golgin B1 AB24568, pChk1 AB47318 (Abcam); PCNA SC7907, pATM SC47739, PML SC5621, AR SC815, chromogranin A SC13090, pATR SC109912, ATR SC1887, Chk1 SC7898 (Santa Cruz Biotechnology); 53BP1 NB100-304 (Novus Biologicals); p53 BD5544147 (Becton Dickinson); Sca1 60032PE (Stem Cell Technologies); SMA A5228, CD49f 60037AD (Millipore-Sigma); and b-actin 4970T (Cell Signaling Technology).
Wu J, & Crowe D.L. (2020). Telomere DNA Damage Signaling Regulates Prostate Cancer Tumorigenesis. Molecular cancer research : MCR, 18(9).
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4

Antibody Characterization for Epigenetic Studies

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Antibodies used in this study were DICER (3363, rabbit; Cell Signaling), ZRF1 (NBP2-12802; Novus Biologicals). RING1B clone (D22F2, 5694, rabbit; Cell Signaling), XPA (GTX103168, mouse; Genetex), XPA (FL-273, rabbit; Santa Cruz) H2B (V119 8135, mouse; Cell Signaling), FLAG (mouse; Sigma), H4K20me2 (39173, rabbit; Active Motif), H4K16ac (39930, rabbit; Active Motif), H3K36me2 (39256, rabbit; Active Motif), CPD (mouse; CosmoBio), HA (C29F4, rabbit; Cell Signaling), XPB (S-19, rabbit; Santa Cruz), MMSET (39880, mouse; Active Motif), 53BP1 (NB100-304, rabbit; Novus), and SETD8 (C18B7, rabbit; Cell Signaling).
Chitale S, & Richly H. (2018). DICER- and MMSET-catalyzed H4K20me2 recruits the nucleotide excision repair factor XPA to DNA damage sites. The Journal of Cell Biology, 217(2), 527-540.
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5

Immunofluorescence Staining of hESCs

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hESCs were seeded onto Matrigel-coated Thermanox Cell Culture Coverslips (Nunc). Cells were washed once and fixed with 4% PFA for 10 min at RT. Cells were then permeabilized with 1% Triton X-100 for 10 min, washed with PBS, and blocked with 3% goat serum in PBS for 1 h at RT. Cells were incubated with primary antibodies diluted in 3% goat serum for 1–2 h at RT and then with secondary antibodies diluted in 3% goat serum for 1 h at RT (Alexa Fluor 488 or Alexa Fluor 568, Molecular Probes). Primary antibodies used included γH2AX (05-636, EMD Millipore), 53BP1 (NB100-304, Novus Biologicals), RAD51 (PC130, EMD Millipore), and OCT4 (19857, Abcam). The nuclei of cells were stained using Hoechst-33342 for 10 min at RT. Coverslips were mounted and imaged using a Carl Zeiss Plan-apochromat 63×/1.4 numerical aperture oil immersion objective on a Carl Zeiss LSM 780 confocal microscope. Images were analyzed using MetaMorph.
Madden-Hennessey K., Gupta D., Radecki A.A., Guild C., Rath A, & Heinen C.D. (2022). Loss of mismatch repair promotes a direct selective advantage in human stem cells. Stem Cell Reports, 17(12), 2661-2673.
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