- PA5-47763 - Invitrogen Antibodies ITGA1 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis from lysates of SH-SY5Y human neuroblastoma cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti- human Integrin a1/CD49a Antigen Affinity-purified Polyclonal Antibody (Product # PA5-47763) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody. A specific band was detected for Integrin a 1/CD49a at approximately 200 kDa (as indicated). This experiment was conducted under reducing conditions.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of CD49a in SH‚SY5Y human neuroblastoma cell line. Samples were incubated in CD49a polyclonal antibody (Product # PA5-47763) using a dilution of 1 µg/mL followed by a HRP-conjugated Anti-Sheep IgG secondary antibody. A specific band was detected for Integrin α 1/CD49a at approximately 200 kDa (as indicated). This experiment was conducted under reducing conditions.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-CD49a Polyclonal Antibody (Product # PA5-47763) and a 200 kDa band corresponding to CD49a was observed across cell lines and tissue tested along with an uncharacterized band at ~40 kDa in MCF7. Membrane Enriched lysates (40 µg lysate) of HeLa (Lane 1), SH-SY5Y (Lane 2), Hep G2 (Lane 3), K-562 (Lane 4), MCF7 (Lane 5), Mouse Kidney (Lane 6), Mouse Mammary gland (Lane 7), Rat Kidney (Lane 8) 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 µg/mL concentration) and detected by chemiluminescence with Rabbit anti-Sheep IgG (H+L Secondary Antibody, HRP (Product # 61-8620, 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 CD49a was performed using 70 % confluent log phase Hep G2 cells. The cells were fixed with 4% paraformaldehyde for 5 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with CD49a Polyclonal Antibody (Product # PA5-47763) at 5 µg/mL concentration in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Sheep IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 (Product # A11015), (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 plasma membrane and cytoplasm localization. Panel e represents low expressing MCF7. Panel f 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 CD49a in immersion fixed paraffin-embedded sections of human stomach cancer tissue. Samples were incubated in CD49a polyclonal antibody (Product # PA5-47763) using a dilution of 10 µg/mL overnight at 4 °C. Before incubation with the primary antibody tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-basic. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown) and counterstained with hematoxylin (blue).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Flow cytometry of CD49a of HeLa human cervical epithelial carcinoma cell line. Samples were incubated in CD49a polyclonal antibody (Product # PA5-47763) or isotype control antibody followed by Phycoerythrin-conjugated Anti-Sheep IgG Secondary Antibody.

PA5-47763