c-SRC is a non-receptor tyrosine kinase, which is activated following stimulation of various cell receptors (E.g. GPCR’s, integrins and receptor tyrosine kinases). This molecule has diverse range of interactions which form a network of channels for intracellular communication, which makes its role difficult to specify. Although, SRC appears to take on a primary role in activating other protein tyrosine kinases. Where receptors dimerise or cluster SRC is recruited, and phosphorylates tyrosine residues within the cytoplasmic domains. As such this gives c-SRC its proto-oncogene status as its elevated expression links with cancer expression as other signals are promoted.

1. HOW DID OUR
ANTIBODY PERFORM?
ANTIBODY CUSTOMER REVIEW: Phospho-c-Src (S75) Polyclonal Antibody (STJ90958)

2. WHAT WAS USED?
Primary antibody: Phospho-c-Src (S75) Polyclonal
Antibody (STJ90958)
Provider: St John's Laboratory
Dilution ratio: 1:500
Application: Western Blot
Materials for validation: LOVO cell line

3. WHAT WAS THE PROTOCOL?
Treatment of materials: Cells were lysed in RIPA buffer (25 mM Tris-HCl
pH 7.6, 150 mM NaCl, 1% Triton X-100, 1% Sodium deoxycholate, 1% SDS,
supplemented with protease inhibitors (PMSF 1mM, Leupeptin 1ug/ml,
Aprotinin 1ug/ml, Pepstatin 1ug/ml). Proteins were reduced and denatured by
heating at 100℃ for 5 min with subsequent cooling before use.
Gel electrophoresis: 60ug/lane
Transfer information: Nitocellulose membrane.
Blocking: Blocked with 5% BSA for 0.5-1hr
Primary Ab Application: Overnight at 4oC, 5% BSA in TBST.
Secondary Ab Application: After washing with TBST 3 times membrane
incubated secondary antibody for 1h at room temperature.
Visualization: ECL kit

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2. Test it in your laboratory using your preferred method.
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