PA5-29349

antibody from Invitrogen Antibodies

Targeting: IDE

Antibody data

Submit

Validation data

IDE protein structure - PA5-29349 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of insulin-degrading enzyme using 30 µg of HepG2 lysate. Samples were loaded onto a 7.5% SDS-PAGE gel and probed with an Insulin-degrading enzyme polyclonal antibody (Product # PA5-29349) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of Insulin-degrading enzyme was achieved by transfecting HeLa with Insulin-degrading enzyme specific siRNAs (Silencer® select Product # s7116, s7118). Western Blot analysis (Fig. a) was performed using Whole cell extracts from the Insulin-degrading enzyme knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with IDE Polyclonal Antibody (Product # PA5-29349, 1:2000 dilution ) 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 Insulin-degrading enzyme.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot was performed using Anti-IDE Polyclonal Antibody (Product # PA5-29349) and a 117 kDa band corresponding to Insulin-degrading enzyme was observed across cell lines and tissue extracts tested. An uncharacterised band (*) was also observed at ~80 kDa. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), Hep G2 (Lane 2), K-562 (Lane 3), A-431 (Lane 4), Caco-2 (Lane 5), U-937 (Lane 6), C2C12 (Lane 7) and tissue extracts (30 µg lysate) of Mouse Liver (Lane 8), Mouse Skeletal Muscle (Lane 9) and Rat Liver (Lane 10) 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 (1:2000 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).