PA5-23402

antibody from Invitrogen Antibodies

Targeting: YWHAZ 14-3-3-zeta, KCIP-1, YWHAD

Immunohistochemistry

Antibody data

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Validation data

Product number: PA5-23402 - Provider product page
Provider: Invitrogen Antibodies
Product name: Anti-14-3-3 zeta Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Reactivity: Human, Mouse, Rat, Bovine, Canine, Porcine
Host: Rabbit
Isotype: IgG
Vial size: 100 µg
Concentration: 0.5 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

YWHAZ protein structure - PA5-23402 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of 14-3-3 zeta in human brain lysate in the 1) absence, 2) presence of immunizing peptide, 3) mouse brain lysate and 4) rat brain lysate using a 14-3-3 zeta polyclonal antibody (Product # PA5-23402) at 4 µg/mL. Goat anti-rabbit Ig HRP secondary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of YWHAZ was achieved by transfecting MCF-7 with YWHAZ specific siRNAs (Silencer® select Product # s14972). Western blot analysis (Fig. a) was performed using modified whole cell extracts from the YWHAZ knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with 14-3-3 zeta Polyclonal Antibody (Product # PA5-23402, 1.5 µg/mL) 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 YWHAZ.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-14-3-3 zeta Polyclonal Antibody (Product # PA5-23402) and a 28 kDa band corresponding to YWHAZ was observed across all the cell lines and tissues tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), A549 (Lane 2), SH-SY5Y (Lane 3), MCF7 (Lane 4), COS-7 (Lane 5), PC-12 (Lane 6), NIH/3T3 (Lane 7), tissue extracts of Mouse Brain (Lane 8) and Rat Brain (Lane 9) were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris 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 (3 µg/mL) 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).