Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [1]
- Immunohistochemistry [1]
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Validation data
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- Product number
- PA5-32638 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Villin Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- Heat-mediated antigen retrieval is recommended prior to staining, using a 10mM citrate buffer, pH 6.0, for 10 minutes followed by cooling at room temperature for 20 min. Following antigen retrieval, incubate samples with primary antibody for 30 min at room temperature. A suggested positive control is small intestine tissue.
- Reactivity
- Human, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 500 µL
- Concentration
- Conc. not determined
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-Villin Polyclonal Antibody (Product # PA5-32638) and a 92kDa band corresponding to Villin was observed along with uncharacterized band (*) across cell line and tissue tested. Whole cell extracts (30 µg lysate) of Caco-2 (Lane 1), COLO 205 (Lane 2), Jurkat (Lane 3) and PANC-1 (Lane 4) as seen in Fig (a).Tissue extracts of Mouse Liver (Lane 1), Mouse Colon (Lane 2) and Mouse Jejunum (Lane 3) as seen in Fig (b) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Relative expression of Villin was observed high in Caco-2 and COLO 205 and low to negative in Jurkat and PANC-1 as seen in Fig (a), similarly expression of Villin was high in Mouse Jejunum and low to negative in Mouse Colon and Mouse Liver as seen in Fig (b).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).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of Villin using anti-Villin Polyclonal Antibody (Product # PA5-32638) in Small Intestine Cancer Tissue. The recommened dilution for this antibody in immunohistochemistry applications is 1:50.