HOW DID OUR ANTIBODY PERFORM?

ANTIBODY CUSTOMER REVIEW: Talin-1 Antibody (STJ97271)

What was used?Primary antibody: STJ97271 Talin-1 Antibody

Provider: St John's Laboratory

Dilution ratio: 1:100

Application: Immunohistochemistry

Materials for validation: Skeletal muscle of human origin (kryo-section)

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What Was The Protocol?Treatment of Materials • Cryo sections were cut into 6um pieces, and left overnight in -80°C

freezer.

Protocol • Primary antibody was incubated overnight at room temperature (23°C).• 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.

What were the results?

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 sarcolemmawhich is in agreement with the predicted subcellular localisation of talin-1, the targetprotein linking integrin proteins to the cytoskeleton. Increased immunoreactivity canbe detected in diseased, degenerating muscle fibres which is in agreement with resultsof in vitro data generated in our laboratory. Unfortunately, antibody generates a markedbackground, also in “healthy” muscle fibres.”

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