HOW DID OUR ANTIBODY PERFORM?

ANTIBODY CUSTOMER REVIEW: Phospho-p70 S6 kinase α (S447) Polyclonal Antibody (STJ90374)

WHAT WAS USED?

Primary Antibody: Phospho-p70 S6 kinase α (S447) Polyclonal Antibody

(STJ90374)

Provider: St John's Laboratory

Dilution ratio: 1:500

Application: Western Blot

Materials for validation: A549 & NIH3T3 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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