- PA5-69083 - Invitrogen Antibodies THOC1 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of Nuclear Matrix Protein p84 was achieved by transfecting HeLa with Nuclear Matrix Protein p84 specific siRNAs (Silencer® select Product # s19396, s19397). Western blot analysis (Fig. a) was performed using Whole cell extracts from the Nuclear Matrix Protein p84 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with Nuclear Matrix Protein p84 Polyclonal Antibody (Product # PA5-69083, 0.75ug/ml ) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution). 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 Nuclear Matrix Protein p84.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-Nuclear Matrix Protein p84 Polyclonal Antibody (Product # PA5-69083) and a 75kDa band corresponding to Nuclear Matrix Protein p84 was observed across cell lines tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), A549 (Lane 2), MDA-MB-231 (Lane 3) and MCF 10A (Lane 4) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). 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 (1ug/ml) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) 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: Immunofluorescence analysis of Nuclear Matrix Protein p84 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 Nuclear Matrix Protein p84 Polyclonal Antibody (Product # PA5-69083) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with,(1:2000 dilution), 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 dilution). 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)
Main image
Experimental details: Immunohistochemical analysis of Nuclear Matrix Protein p84 in paraffin-embedded human kidney (epithelial cells of the renal tubule) tissue. Sample was probed with a Nuclear Matrix Protein p84 polyclonal antibody (Product # PA5-69083) at a dilution of 4.0-8.0 µg/mL. Images were taken at 400x magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of Nuclear Matrix Protein p84 in paraffin-embedded human kidney (epithelial cells of the renal tubule) tissue. Sample was probed with a Nuclear Matrix Protein p84 polyclonal antibody (Product # PA5-69083) at a dilution of 4.0-8.0 µg/mL. Images were taken at 400x magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of Nuclear Matrix Protein p84 in paraffin-embedded human kidney (epithelial cells of the renal tubule) tissue. Sample was probed with a Nuclear Matrix Protein p84 polyclonal antibody (Product # PA5-69083) at a dilution of 4.0-8.0 µg/mL. Images were taken at 400x magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of Nuclear Matrix Protein p84 in paraffin-embedded human kidney (epithelial cells of the renal tubule) tissue. Sample was probed with a Nuclear Matrix Protein p84 polyclonal antibody (Product # PA5-69083) at a dilution of 4.0-8.0 µg/mL. Images were taken at 400x magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of Nuclear Matrix Protein p84 in paraffin-embedded human kidney (epithelial cells of the renal tubule) tissue. Sample was probed with a Nuclear Matrix Protein p84 polyclonal antibody (Product # PA5-69083) at a dilution of 4.0-8.0 µg/mL. Images were taken at 400x magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of Nuclear Matrix Protein p84 in paraffin-embedded human kidney (epithelial cells of the renal tubule) tissue. Sample was probed with a Nuclear Matrix Protein p84 polyclonal antibody (Product # PA5-69083) at a dilution of 4.0-8.0 µg/mL. Images were taken at 400x magnification.

PA5-69083