MA5-15006

antibody from Invitrogen Antibodies

Targeting: RICTOR AVO3, KIAA1999, MGC39830, PIA

Antibody data

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Validation data

RICTOR protein structure - MA5-15006 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-RICTOR Polyclonal Antibody (Product # MA5-15006) and a 192 kDa band corresponding to RICTOR was observed across cell lines. Whole cell extracts (40 µg lysate) of SW480 (Lane 1), BeWo (Lane 2), HeLa (Lane 3), MOLT4 (Lane 4), HCT 116 serum starved overnight (Lane 5), HCT116 serum starved overnight and treated with EGF (20 ng/mL for 45 mins) (Lane 6), HCT116 serum starved overnight, treated with Curcumin 25 µM for 2 hrs and then treated with EGF (20 ng/mL for 45 mins) (Lane 7), A549 (Lane 8), A549 treated with MG132 5 µM for 3 hrs (Lane 9) 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 (*1:1000) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20000 dilution) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Atto Ultimate Sensitivity Substrate (Product # A38556).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of RICTOR was achieved by transfecting HeLa with RICTOR specific siRNAs (Silencer® select Product # s48408, S226001). Western blot analysis (Fig. a) was performed using Whole cell extracts from the RICTOR knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with RICTOR Polyclonal Antibody (Product # MA5-15006, 1:1000) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20000 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 RICTOR.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot was performed using Anti-RICTOR Monoclonal Antibody (J.644.6) (Product # MA5-15006) and a 200 kDa band corresponding to Rapamycin-insensitive companion of mTOR was observed in A549 cells treated with MG132 (5 µM for 4hr). Freshly prepared Whole cell extracts (30 µg lysate) of A549 (Lane 1), A549 treated with MG132 (5 µM for 4hr) (Lane 2) 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 (1:1000 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).