PA5-17589

antibody from Invitrogen Antibodies

Targeting: PRKCZ PKC2

Antibody data

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Validation data

PRKCZ protein structure - PA5-17589 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of PKC zeta was achieved by transfecting HeLa with PKC zeta specific siRNAs (Silencer® select Product # s11128). Western blot analysis (Fig. a) was performed using whole cell extracts from the PKC zeta knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with PKC zeta Polyclonal Antibody (Product # PA5-17589, 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 PKC zeta.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-PKC zeta Polyclonal Antibody (Product # PA5-17589) and a 67 kDa, corresponding to PKC zeta was observed in all cell lysates tested. Whole cell lysates (30 µg lysate) of HEK-293 (Lane 1), HeLa (Lane 2), Hep G2 (Lane 3), MDA-MB-231 (Lane 4) and THP-1 (Lane 5) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris 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). An uncharacterized band (*) was also observed at ~35 kDa.