MA5-14909

antibody from Invitrogen Antibodies

Targeting: SHH HHG1, HLP3, HPE3, MCOPCB5, SMMCI, TPT, TPTPS

Antibody data

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Validation data

SHH protein structure - MA5-14909 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of Sonic hedgehog protein was achieved by transfecting A549 with Sonic hedgehog protein specific siRNAs (Silencer® select Product # S12818, S12817). Western Blot analysis (Fig. a) was performed using Whole cell extracts from the Sonic hedgehog protein knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with SHH Monoclonal Antibody (G.162.10) (Product # MA5-14909, 1:1000 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 Sonic hedgehog protein.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot was performed using Anti-SHH Monoclonal Antibody (G.162.10) (Product # MA5-14909) and a ~50 kDa band corresponding to Sonic hedgehog protein was observed across cell lines tested. Whole cell extracts (30 µg lysate) of A549 (Lane 1), PANC-1 (Lane 2), MIA PaCa-2 (Lane 3), Hep G2 (Lane 4), U-2 OS (Lane 5) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # LC2001) 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 SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).