PA5-17237

antibody from Invitrogen Antibodies

Targeting: WWOX FOR, SDR41C1, WOX1

Antibody data

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Validation data

WWOX protein structure - PA5-17237 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 1 WWOX expression is reduced in canine MCTs and malignant mast cell lines. RNA was collected from (a) normal canine BMCMCs and fresh frozen canine MCT tissues and (b) malignant canine BR and C2 mast cell lines and quantitative RT-PCR for WWOX was performed. c Canine BR and C2 mast cell lines were left untreated or incubated with 100 ng/mL rcSCF for 24 h prior to collection. Protein lysates were generated, separated by SDS-PAGE and Western blotting for WWOX and beta-actin was performed. d Fresh frozen canine MCT tissues and e malignant canine (BR and C2) and mouse (C57 and P815) mast cell lines were processed for protein lysates. Protein was separated by SDS-PAGE and Western blotting for WWOX and beta-actin was performed. Full-length blots are presented in Supplementary Fig. 1 . Three independent experiments were performed and all reactions were performed in triplicate. The experiments were repeated 3 times in the cell lines and twice for normal canine BMCMCs

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 2 Generation of canine mast cell lines expressing WWOX lentiviral constructs or shRNAs targeting WWOX. a Full-length canine WWOX cDNA was PCR amplified from canine testes and cloned into the pCDH-copGFP lentiviral expression plasmid (Systems Biosciences). Canine BR cells transduced with empty control vector (EV) or WWOX lentivirus were sorted 72 h post-transduction to greater than 95% purity based on GFP expression. WWOX expression was assessed by qRT-PCR and Western blotting in empty vector or WWOX-expressing BR cells. b Canine C2 cells transduced with either scramble (Scr) control vector or pGreenPuro-shWWOX-copGFP lentiviral constructs (shWWOX-554 or shWWOX-1304) underwent FACs-mediated cell sorting based on GFP expression 72 h post-transduction. RNA and protein lysates were generated and qRT-PCR and Western blotting for WWOX and beta-actin was performed to confirm efficiency of WWOX knockdown. Full-length blots are presented in Supplementary Fig. 1 . Gene expression was calculated using the DeltaCt method and all samples were normalized to 18S. Data represent mean +/- SD of three independent experiments and representative blots from three experimental replicates are shown. (Bars: SD. Statistical analysis: one-way ANOVA, * p