PA5-30462

antibody from Invitrogen Antibodies

Targeting: ATG4B APG4B, AUTL1, DKFZp586D1822, KIAA0943

Western blot (Supportive data in Antibodypedia)

Immunohistochemistry (Data presented on provider website)

Antibody Data

Product number

PA5-30462

Provider

Invitrogen Antibodies

Product name

Anti-ATG4B Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant protein fragment

Description

Recommended positive controls: HepG2, HCT116. Predicted reactivity: Human (99%), Mouse (91%), Rat (91%), Pig (91%), Bovine (86%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Reactivity

Human

Host

Rabbit

Isotype

IgG

Vial size

100 µL

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 ATG4B using 30 µg of HepG2 lysate. Samples were loaded onto a 10% SDS-PAGE gel and probed with an ATG4B polyclonal antibody (Product # PA5-30462) at a dilution of 1:1000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of ATG4B was achieved by transfecting MCF7 with ATG4B specific siRNAs (Silencer® select Product # s23244, s23246). Western blot analysis (Fig. a) was performed using whole cell extracts from the ATG4B knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with ATG4B Polyclonal Antibody (Product # PA5-30462, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 0.25µg/ml, 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 ATG4B.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-ATG4B Polyclonal Antibody (Product # PA5-30462) and a 45 kDa band corresponding to ATG4B was observed across cell lines along with an uncharacterized band (*) at ~80 kDa in the cell lines tested. Whole cell extracts (30 µg lysate) of MCF7 (Lane 1), Jurkat (Lane 2), PC-3 (Lane 3) and Hep G2 (Lane 4) were electrophoresed using 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).