- PA1-16566 - Invitrogen Antibodies TP53BP1 antibody

Submitted references Loss of CENP-I Impairs Homologous Recombination and Sensitizes Cells to PARP1 Inhibition.

Dang TT, Morales JC
Cancers 2021 Jun 26;13(13)

Multi-omics analyses of radiation survivors identify radioprotective microbes and metabolites.

Guo H, Chou WC, Lai Y, Liang K, Tam JW, Brickey WJ, Chen L, Montgomery ND, Li X, Bohannon LM, Sung AD, Chao NJ, Peled JU, Gomes ALC, van den Brink MRM, French MJ, Macintyre AN, Sempowski GD, Tan X, Sartor RB, Lu K, Ting JPY
Science (New York, N.Y.) 2020 Oct 30;370(6516)

Involvement of POLA2 in Double Strand Break Repair and Genotoxic Stress.

Dang TT, Morales JC
International journal of molecular sciences 2020 Jun 15;21(12)

Increased DNA double-strand break was associated with downregulation of repair and upregulation of apoptotic factors in rat hippocampus after alcohol exposure.

Suman S, Kumar S, N'Gouemo P, Datta K
Alcohol (Fayetteville, N.Y.) 2016 Aug;54:45-50

Exposure to heavy ion radiation induces persistent oxidative stress in mouse intestine.

Datta K, Suman S, Kallakury BV, Fornace AJ Jr
PloS one 2012;7(8):e42224

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot analysis of human 53BP1, using Product # PA1-16566. Sample: Whole cell lysate (20 µg/lane) from U2Os cells resolved on a 3 to 8% trisacetate gel.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of 53BP1 in 20 µg/lane whole cell lysate from U2Os cells. Samples were incubated in 53BP1 polyclonal antibody (Product # PA1-16566). Resolved on a 3 to 8% trisacetate gel. A band appears at approximately the theoretical molecular weight of 214 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of TP53-binding protein 1 was achieved by transfecting HeLa cells with TP53-binding protein 1 specific siRNAs (Silencer® select Product # s14315, s14314). Western blot analysis (Fig. a) was performed using Nuclear enriched extracts from the TP53-binding protein 1 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with 53BP1 Polyclonal Antibody (Product # PA1-16566, 1:5000 ) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to TP53-binding protein 1.Pick up in positive cell lines is observed as a streak like pattern which is expected of 53BP1 target.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-53BP1 Polyclonal Antibody (Product # PA1-16566) and a ~270kDa band corresponding to TP53-binding protein 1 was observed across cell lines tested . Nuclear enriched extracts (30 µg lysate) of HeLa (Lane 1), A549 (Lane 2), MDA-MB-231 (Lane 3), MDA-MB-231 (Lane 4), MCF7 (Lane 5), U-2 OS (Lane 6), K-562 (Lane 7), U-87 MG (Lane 8) were electrophoresed using NuPAGE™ 3-8% Tris-Acetate Protein Gel (Product # EA0378BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # LC2001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:5000) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).53BP1 picks up in a streak pattern as observed in the positive cell lines here.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of TP53-binding protein 1 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with 53BP1 Polyclonal Antibody (Product # PA1-16566) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical analysis of 53BP1 in placental villi. Samples were incubated in 53BP1 polyclonal antibody (Product # PA1-16566). Antibody at 40X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Flow cytometry of 53BP1 in Ntera2 cells (blue) and a matched isotype control (orange). Samples were incubated in 53BP1 polyclonal antibody (Product # PA1-16566) using a dilution of 1.0 µg/mL for 30 minutes at room temperature followed by a Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight™ 550 (Product # SA5-10033). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry of 53BP1 in HeLa cells (blue) and a matched isotype control (orange). Samples were incubated in 53BP1 polyclonal antibody (Product # PA1-16566) using a dilution of 5 µg/mL for 30 minutes at room temperature. Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Both antibodies were conjugated to Alexa Fluor 488.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 6 Assessing 53BP1 foci and cell death in intestinal sections after gamma and 56 Fe radiation exposure. A) Intestinal sections from 6 to 8 week control, 1-year post-exposure control, 2 Gy gamma, and 1.6 Gy 56 Fe 1-year post-exposure mice were immunofluorescently stained for 53BP1. B) Quantification of 53BP1 foci in intestinal cell nuclei counterstained with DAPI. C) Quantification of TUNEL staining of intestinal sections after exposure to gamma and 56 Fe radiation. *significant compared to 6-8 wks unirradiated control; **significant compared to 1-year post-exposure unirradiated control; ***significant compared to gamma radiation.

PA1-16566

Antibody data