For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 10
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro215 of human OCA-T1 protein. Antibodies are purified by peptide affinity chromatography.
OCA-T1 is encoded by the gene POU2AF2 (C11orf53), which was identified in a single-cell RNA sequencing study to be expressed selectively in tuft cells from mouse and human tissues (1). The expression of OCA-T1 was observed to occur in a pattern strikingly similar to that of POU2F3, a master regulator of tuft cell development whose expression also defines a tuft cell-like variant (SCLC-P) of small cell lung cancers (2). Although containing no recognizable structural domains, OCA-T1 showed minor sequence similarity with OCA-B, a protein encoded by the POU2AF1 gene and reported to be an activator of selected Class II POU domain transcription factors (3). Functional studies revealed that OCA-T1 forms a protein complex with POU2F3; these proteins furthermore share genomic binding sites and promote the expression of genes directly associated with tuft cell development. Lastly, Crispr/Cas9-mediated deletion of POU2AF2 in mice revealed that OCA-T1 is functionally required for normal tuft cell development (1). Collectively, these findings suggest that OCA-T1 is a critical component of the POU2F3-containing transcriptional complex that regulates the expression of genes governing tuft cell development.
Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.
Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.