Akt1 (a Serine/Threonine Protein Kinase) is involved in cell survival signal pathways. In response to PDGF (platelet-derived growth factor) Akt rapidly signals through its Serine/Threonine kinase by phosphorylating components that upregulate protein synthesis, prevent apoptosis and stimulate cell survival.

1. HOW DID OUR
ANTIBODY PERFORM?
CUSTOMER REVIEW: Phospho-Akt1 (T450) Polyclonal Antibody (STJ90168)

2. WHAT WAS USED?
Primary Antibody: Phospho-Akt1 (T450) Polyclonal
Antibody (STJ90168)
Provider: St John's Laboratory
Dilution ratio: 1:500
Application: Western Blot
Materials for validation: C2C12 & RD cells

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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