- PA5-29068 - Invitrogen Antibodies COL6A1 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of COL6A1 using 50 µg of mouse brain lysate. Samples were loaded onto a 5% SDS-PAGE gel and probed with a COL6A1 polyclonal antibody (Product # PA5-29068) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot analysis of COL6A1 was performed by separating 30 µg of Whole cell extracts by 7.5% SDS-PAGE. Proteins were transferred to a membrane and probed with a COL6A1 Polyclonal Antibody (Product # PA5-29068) at a dilution of 1:500. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-COL6A1 Polyclonal Antibody (Product # PA5-29068) and a 140 kDa band corresponding to COL6A1 was observed in IMR-90 and was absent in SK-O-V3. Whole cell extracts (30 µg lysate) of IMR-90 (Lane 1) and SK-O-V3 (Lane 2) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). 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 (1:1000 dilution) 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: Immunocytochemistry-Immunofluorescence analysis of COL6A1 was performed in DIV10 rat E18 primary cortical neuron cells fixed in 4% paraformaldehyde at RT for 15 min. Green: COL6A1 Polyclonal Antibody (Product # PA5-29068) diluted at 1:500. Red: beta Tubulin 3/ Tuj1, stained by beta Tubulin 3/ Tuj1 antibody. Blue: Fluoroshield with DAPI.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of COL6A1 was performed using 70% confluent log phase IMR-90 and SK-O-V3 cells. The cells were fixed with 4% paraformaldehyde for 5 minutes, permeabilized with 0.1% Triton™ X-100 for 5 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with COL6A1 Polyclonal Antibody (Product # PA5-29068) 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 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). Panel d represents the merged image showing cytosolic localization. Panel (e-h) represents SK-O-V3 cells having no expression of COL6A1. The images were captured at 60 magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: COL6A1 Polyclonal Antibody detects COL6A1 protein at cytosol on human hepatoma by immunohistochemical analysis. Sample: Paraffin-embedded hepatoma. COL6A1 Polyclonal Antibody (Product # PA5-29068) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: COL6A1 Polyclonal Antibody detects COL6A1 protein at cytosol on mouse liver by immunohistochemical analysis. Sample: Paraffin-embedded mouse liver. COL6A1 Polyclonal Antibody (Product # PA5-29068) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

PA5-29068