PA5-54425
antibody from Invitrogen Antibodies
Targeting: WIPI2
ATG18B, ATG21, CGI-50, DKFZP434J154, DKFZp686P02188, FLJ12979, FLJ14217, FLJ42984
Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
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Validation data
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- 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.
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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).
- 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.
- 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).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent staining of WIPI2 in human cell line A-431 shows positivity in nucleus but excluded from the nucleoli. Samples were probed using a WIPI2 Polyclonal Antibody (Product # PA5-54425).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical staining of WIPI2 in human testis using a WIPI2 Polyclonal Antibody (Product # PA5-54425) shows cytoplasmic positivity in cells in seminiferous ducts.