PA1-28295

antibody from Invitrogen Antibodies

Targeting: CDC42 CDC42Hs, G25K

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

PA1-28295

Provider

Invitrogen Antibodies

Product name

Anti-Cdc42 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Other

Description

PA1-28295 detects CDC42 from human, mouse, rat samples.

Reactivity

Human, Mouse, Rat

Host

Chicken/Avian

Isotype

IgY

Vial size

50 µg

Concentration

1 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Cdc42 in E coli-derived fusion protein using a Cdc42 polyclonal antibody (Product # PA1-28295) at a dilution of 1:2000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

KD of Cdc42 was achieved by transfecting A-431 with Cdc42 specific siRNAs (Silencer® select Product # s2765, s2766). Western blot analysis (Fig. a) was performed using whole cell extracts from the Cdc42 KD cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with Cdc42 Polyclonal Antibody (Product # PA1-28295, 1:1000 dilution) and Goat anti-Chicken IgY (H+L) Secondary Antibody, HRP (Product # A16054, 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 Cdc42. An uncharacterized band at ~80 kDa was observed in the samples tested..

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-Cdc42 Polyclonal Antibody (Product # PA1-28295) and 21 kDa band corresponding to Cdc42 was observed across cell lines and tissues tested along with an uncharacterized band at ~80 kDa. Whole cell extracts (30 µg lysate) of U-937 (Lane 1), A-431 (Lane 2), T-47D (Lane 3), K-562 (Lane 4), HeLa (Lane 5), Hep G2 (Lane 6), tissue extracts of Mouse Brain (Lane 7) and Rat Brain (Lane 8) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0342BOX). 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 Goat anti-Chicken IgY (H+L) Secondary Antibody, HRP (Product # A16054, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).