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Syndecan-4 is a member of the Syndecan proteoglycan family and is encoded by the SDC4 gene in humans. It is a cell surface proteoglycan that is known to regulate exosome biogenesis. They are usually present on the plasma membrane of epithelial and fibroblast cells. The Syndecan-4 antibody detects endogenous levels of Syndecan-4 protein.
1. HOW DID OUR ANTIBODY PERFORM? ANTIBODY CUSTOMER REVIEW: Syndecan-4 Antibody (STJ95856)
2. What was used? Primary antibody: STJ95856 Syndecan-4 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 ¡°Immunohistochemical investigation showed immunoreactivity at the sarcolemma which is in perfect accordance with the predicted subcellular localisation of syndecan- 4, the target protein. Increased immunoreactivity can be detected in diseased, degenerating muscle fibres which is in agreement with results of in vitro data generated in our laboratory. Antibody does not seem to generate a marked background..¡±
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