- PA5-17962 - Invitrogen Antibodies SET antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of SET by a SET monoclonal antibody (Product # PA5-17962) at a concentration of 0.3 µg/mL. Human Kidney lysate (35µg protein in RIPA buffer). Primary incubation was 1 hour. Detected by chemiluminescence.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot staining of Human Kidney extracts using Product # PA5-17962 at a concentration of 2 µg/mL, the primary antibody incubation was 1 hour and the detection method was chemiluminescence.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of SET by a SET monoclonal antibody (Product # PA5-17962) at a concentration of 0.01 µg/mL. Daudi (A) and Molt4 (B) cell lysate (35µg protein in RIPA buffer). Primary incubation was 1 hour. Detected by chemiluminescence.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of SET was achieved by transfecting HeLa with SET specific siRNAs (Silencer® select Product # s12704, s12705). Western blot analysis (Fig. a) was performed using whole cell extracts from the SET knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with SET Polyclonal Antibody (Product # PA5-17962, 0.3 µg/mL) and Rabbit anti-Goat IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27014, 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 SET.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-SET Polyclonal Antibody (Product # PA5-17962) and a 38 kDa corresponding to SET was observed across cell lines tested except MCF 10A. The expression of SET is low in MCF 10A when compared to MCF-7 and MDA-MB-231 as it is overexpressed in breast cancers. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), MCF-7 (Lane 2), MDA-MB-231 (Lane 3), MOLT-4 (Lane 4), Jurkat (Lane 5) and MCF 10A (Lane 6) 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 (0.3 µg/mL) and detected by chemiluminescence with Rabbit anti-Goat IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27014, 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: Immunohistochemical analysis of SET in Human Adrenal Gland using a SET monoclonal antibody (Product #PA5-17962) at 3.75 µg/mL. The Human Adrenal Gland tissue section was paraffin embeded and detected using steamed antigen retrieval with citrate buffer pH 6, AP-staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical analysis of SET in Human Testis using a SET monoclonal antibody (Product #PA5-17962) at 3.75 µg/mL. The Human Testis tissue section was paraffin embeded and detected using steamed antigen retrieval with citrate buffer pH 6, AP-staining.

PA5-17962