- PA5-78523 - Invitrogen Antibodies RIF1 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of RIF1 in whole cell lysate using 30 µg of protein. Samples were separated with 5% SDS-PAGE and incubated with RIF1 polyclonal antibody (Product # PA5-78523) using a dilution of 1:2000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: RIF1 Polyclonal Antibody detects Rif1 protein by western blot analysis. Various whole cell extracts (30 µg) were separated by 5% SDS-PAGE, and the membrane was blotted with RIF1 Polyclonal Antibody (Product # PA5-78523) diluted at 1:2,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: RIF1 Polyclonal Antibody detects Rif1 protein by western blot analysis. Various whole cell extracts (30 µg) were separated by 5% SDS-PAGE, and the membrane was blotted with RIF1 Polyclonal Antibody (Product # PA5-78523) diluted at 1:2,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of Telomere-associated protein RIF1 was achieved by transfecting K-562 with Telomere-associated protein RIF1 specific siRNAs (Silencer® select Product # S30376, S30378). Western blot analysis (Fig. a) was performed using Nuclear enriched extracts from the Telomere-associated protein RIF1 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with RIF1 Polyclonal Antibody (Product # PA5-78523, 1:2000 ) 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 Telomere-associated protein RIF1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-RIF1 Polyclonal Antibody (Product # PA5-78523) and a 274kDa band corresponding to Telomere-associated protein RIF1 was observed. Nuclear enriched extracts (40 µg lysate) of K-562 (Lane 1), HEK-293 (Lane 2) and MCF7 (Lane 3) were electrophoresed using NuPAGE™ 3-8% Tris-Acetate Protein Gel (Product # EA0378BOX). 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 (1:2000) 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: RIF1 Polyclonal Antibody detects Rif1 protein at nucleus by immunofluorescent analysis. Sample: HeLa cells were fixed in 4% paraformaldehyde at RT for 15 min. Green: Rif1 protein stained by RIF1 Polyclonal Antibody (Product # PA5-78523) diluted at 1:500. Blue: Hoechst 33342 staining. Scale bar = 10 µm.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of Telomere-associated protein RIF1 was performed using 70% confluent log phase NIH:OVCAR-3 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 RIF1 Polyclonal Antibody (Product # PA5-78523) at 1:200 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: Rif1 antibody detects Rif1 protein at cytoplasm and nucleus in human breast carcinoma by immunohistochemical analysis. Sample: Paraffin-embedded human breast carcinoma. Rif1 antibody (Product # PA5-78523) diluted at 1:500. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Rif1 antibody detects Rif1 protein at cytoplasm and nucleus in human cervical carcinoma by immunohistochemical analysis. Sample: Paraffin-embedded human cervical carcinoma. Rif1 antibody (Product # PA5-78523) diluted at 1:500. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

PA5-78523