MA5-27619

antibody from Invitrogen Antibodies

Targeting: HIF1AN DKFZp762F1811, FIH1, FLJ20615, FLJ22027

Immunocytochemistry

Immunohistochemistry

Antibody data

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Validation data

HIF1AN protein structure - MA5-27619 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of HIF1AN in human cell lysates with 15 µg of sample. The sample was blocked with 1.5% BSA, incubated with HIF1AN monoclonal antibody (Product # MA5-27619) using a dilution of 1:1000 (2 hours at 4°C), followed by Sheep Anti-Mouse HRP (1 hour at RT).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of HIF1AN was achieved by transfecting HCT 116 with HIF1AN specific siRNAs (Silencer® select Product # s31198, s31197). Western blot analysis (Fig. a) was performed using whole cell extracts from the HIF1AN knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with HIF1AN Monoclonal Antibody (FIH 162c) (Product # MA5-27619, 1:500 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 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 HIF1AN.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-HIF1AN Monoclonal Antibody (FIH 162c) (Product # MA5-27619) and a band at 40 kDa corresponding to HIF1AN was observed across the cell lines tested. Modified whole cell extracts (1% SDS) (30 µg lysate) of HeLa (Lane 1), Jurkat (Lane 2), MCF7 (Lane 3), HCT 116 (Lane 4), LNCaP (Lane 5), DU 145 (Lane 6), HEK-293 (Lane 7) and K-562 (Lane 8) were electrophoresed using Novex® NuPAGE® 4-12% % Bis-Tris gel (Product # NP0321BOX). 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 (1:500 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).