PA5-54425

antibody from Invitrogen Antibodies

Targeting: WIPI2 ATG18B, ATG21, CGI-50, DKFZP434J154, DKFZp686P02188, FLJ12979, FLJ14217, FLJ42984

Antibody data

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Validation data

Product number: PA5-54425 - Provider product page
Provider: Invitrogen Antibodies
Product name: WIPI2 Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Recombinant full-length protein
Description: Immunogen sequence: MKVLHTIRET PPNPAGLCAL SINNDNCYLA YPGSATIGEV QVFDTINLRA ANMIPAHDSP LAALAFDASG TKLATASEKG TVIRVF Highest antigen sequence identity to the following orthologs: Mouse - 99%, Rat - 100%.
Reactivity: Human
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Concentration: 0.2 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

WIPI2 protein structure - PA5-54425 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of WIPI2 in control (vector only transfected HEK293T lysate) and WIPI2 over-expression lysate (Co-expressed with a C-terminal myc-DDK tag (~3.1 kDa) in mammalian HEK293T cells). Samples were probed using a WIPI2 Polyclonal Antibody (Product # PA5-54425).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: KD of WIPI2 was achieved by transfecting SH-SY5Y with WIPI2 specific siRNAs (Silencer® select Product # S25100, S25101). Western blot analysis (Fig. a) was performed using membrane enriched cell extracts from the WIPI2 KD cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with WIPI2 Polyclonal Antibody (Product # PA5-54425, 0.4 µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (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 WIPI2. An uncharacterized band (*) was observed at ~70 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-WIPI2 Rabbit Polyclonal Antibody (Product # PA5-54425) and a 49 kDa band corresponding to WIPI2 was observed across cell lines tested along with an uncharacterized band (*) at ~70 kDa. Membrane enriched cell extracts (30 µg lysate) of A-431 (Lane 1), THP-1 (Lane 2), Jurkat (Lane 3), SH-SY5Y (Lane 4), Raji (Lane 5) and K-562 (Lane 6) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX). 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 (0.4 µg/mL) and detected by chemiluminescence Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (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).