Immunofluorescence Antibody Customer Review for Laminin α-1 Polyclonal Antibody (STJ93888)
Antibody Customer Review for Phospho-p70 S6 Kinase a (S418) Polyclonal Antibody (STJ91046)
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Transcript of Antibody Customer Review for Phospho-p70 S6 Kinase a (S418) Polyclonal Antibody (STJ91046)
HOW DID OUR ANTIBODY PERFORM?
ANTIBODY CUSTOMER REVIEW: Phospho-p70 S6 kinase α (S418) Polyclonal Antibody (STJ91046)
WHAT WAS USED?
Primary antibody: Phospho-p70 S6 kinase α (S418) antibody (STJ91046)
Provider: St John's Laboratory
Dilution ratio: 1:500
Application: Western Blot
Materials for validation: HCT116 cells
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/laneTransfer information: Nitocellulose membrane.Blocking: Blocked with 5% BSA for 0.5-1hrPrimary 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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