- PA5-34802 - Invitrogen Antibodies PARP1 antibody

Submitted references Dissecting the molecular determinants of clinical PARP1 inhibitor selectivity for tankyrase1.

Ryan K, Bolaňos B, Smith M, Palde PB, Cuenca PD, VanArsdale TL, Niessen S, Zhang L, Behenna D, Ornelas MA, Tran KT, Kaiser S, Lum L, Stewart A, Gajiwala KS
The Journal of biological chemistry 2021 Jan-Jun;296:100251

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of PARP1 using 30 µg of A549 lysate. Samples were loaded onto a 7.5% SDS-PAGE gel and probed with a PARP1 polyclonal antibody (Product # PA5-34802) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot analysis of PARP1 was performed by separating 30 µg of non-transfected (–) and transfected (+) 293T whole cell extracts by 5% SDS-PAGE. Proteins were transferred to a membrane and probed with a PARP1 Polyclonal Antibody (Product # PA5-34802) at a dilution of 1:5000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of PARP1 was performed by separating 30 µg of various whole cell extracts by 5% SDS-PAGE. Proteins were transferred to a membrane and probed with a PARP1 Polyclonal Antibody (Product # PA5-34802) at a dilution of 1:1000 and a HRP-conjugated anti-rabbit IgG secondary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of PARP was achieved by transfecting HEK-293 with PARP specific siRNAs (Silencer® select Product # S1098, S1099). Western blot analysis (Fig. a) was performed using Nuclear enriched extracts from the PARP knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with PARP Polyclonal Antibody (Product # PA5-34802, 1:1000 ) 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 PARP.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-PARP Polyclonal Antibody(Product # PA5-34802) and a 116kDa band corresponding to PARP was observed across all tested cell lines. Nuclear enriched extracts (30 µg lysate) of Jurkat (Lane 1), HeLa (Lane 2), MDA-MB-231 (Lane 3), HEK-293 (Lane 4), PC-12 (Lane 5) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000) 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).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry-Immunofluorescence analysis of PARP1 was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: PARP1 Polyclonal Antibody (Product # PA5-34802) diluted at 1:500. Red: phalloidin, a cytoskeleton marker. Scale bar = 10 µm.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry-Immunofluorescence analysis of PARP1 was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: PARP1 Polyclonal Antibody (Product # PA5-34802) diluted at 1:500. Red: phalloidin, a cytoskeleton marker. Scale bar = 10 µm.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical analysis of paraffin-embedded HeLa xenograft, using PARP1 antibody [N1N2], N-term (Product # PA5-34802) antibody at 1:500 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: ChIP assay analysis of PARP1 was performed in HeLa chromatin extract using 5 µg of either normal rabbit IgG or PARP1 Polyclonal Antibody (Product # PA5-34802). The precipitated DNA was detected by PCR with primer set targeting to HSP70.1 promoter.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Cross-linked ChIP was performed with Raji chromatin extract and 5 µg of either control rabbit IgG or PARP1 Polyclonal Antibody (Product # PA5-34802). The precipitated DNA was detected by PCR with primer set targeting to S100A9 promoter.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: PARP1 antibody [N1N2], N-term immunoprecipitates PARP1 protein in IP experiments. IP samples: HCT-116 whole cell extract. A. 30 µg HCT-116 whole cell extract. B. Control with 4 µg of preimmune Rabbit IgG. C. Immunoprecipitation of PARP1 protein by 4 µg PARP1 antibody [N1N2], N-term (Product # PA5-34802). 5 % SDS-PAGE. The immunoprecipitated PARP1 protein was detected by PARP1 antibody [N1N2], N-term (Product # PA5-34802) diluted at 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 1 Talazoparib binds to TNKS1 at a clinically relevant concentration in cultured cells. NCI-H1048 cell lysate was pretreated with indicated concentrations of talazoparib or olaparib (30 min) prior to the addition of talazoparib- or olaparib-biotin probes (30 min). Proteins bound to streptavidin were released and subjected to Western blot analysis for either PARP1 ( top ) or TNKS1 ( bottom ). Relative quantitation (Rel Quant) was determined using ImageJ and is displayed below the plots. PARP1, poly(ADP-ribosyl) polymerase 1; TNKS1, tankyrase1.

PA5-34802

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