Antibody Customer Review for Radixin Polyclonal Antibody (STJ95338)

Target information

Radixin is a cytoskeletal protein that is coded by the RSX gene in humans. It is part of the ERM protein family along with ezrin and moesin. It is thought to play a significant role in the binding of the barbed end of actin filaments to the plasma membrane. The Radixin antibody detects endogenous levels of the radixin protein.

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2. What was used? Primary antibody: STJ95338 Radixin Antibody Provider: St John’s Laboratory Dilution ratio: 1:100 Application: Immunohistochemistry Materials for validation: Skeletal muscle of human origin (kryo-section) Click for product data sheet PDF
3. What Was The Protocol? Treatment of Materials ? Cryo sections were cut into 6um pieces, and left overnight in -80¡ãC freezer. Protocol ? Primary antibody was incubated overnight at room temperature (23¡ãC). ? The sample was washed for 2 hours with multiple changes of PBS. ? Amplification occurred through NOVOLINK Leica microsystems, then the primary antibody was incubated for 1 hour. ? The sample was then washed again for 1 hour with at least 3 changes of PBS. ? The secondary antibody was incubated for 30 minutes and then washed for 2 hours using PBS with multiple changes. ? The sample was then stained for 2 minutes using DAB. ? The sample was counterstained for 3 seconds using Harris¡¯ haematoxylin and then washed with tap water for 2 minutes until blueing. ? The sample was then dehydrated and mounted in entellan.
4. What were the results?
5. What did the customer think? Antibody Specificity: Antibody RPPA Rating: Testing results were provided independently by A. Roos at Newcastle University. They stated ¡°The highly conserved radixin functions as molecular linker between the actin cytoskeleton and transmembrane receptors. Thus increased subsarcolemmal immunoreactivity in a proportion of myofibres is in accordance with this function and the predicted subcellular localization. Remarkably, expression seems to be fibre-type specific. We will perform further (co)staining to confirm this assumption. Moreover, full-length radixin is known to be able to localise to the nucleus. In this context, increased immunoreactivity in (pathological) centralized myonuclei might be a specific finding. Focal increase in degenerating myofibres is in accordance with our proteomic in vitro data. Antibody does not seem to generate a marked background.¡±
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