Bicinchoninic acid (BCA) protein assay (Pierce Biotechnology, Rockford, IL, USA) was used to determine the protein content in each sample. Quantified each sample concentration to 60-80 µg and proteins were fractionated on SDS-PAGE, transferred onto Hybond enhanced chemiluminescence nitrocellulose membranes (Amersham, Little Chalfont, UK) and detected with antibodies against P-gp (GeneTex, Inc. Irvine, CA, USA), the mammalian target of rapamycin (mTOR) (Cell Signaling, Danvers, MA, USA), phosph-mTOR (Cell Signaling), protein kinase B (AKT) (Santa Cruz Biotechnology, Inc. Santa Cruz, CA, USA), phosph-AKT (Santa Cruz Biotechnology, Inc.), p70s6K (Cell Signaling), phosph-p70s6K (Cell Signaling), caspase 3 (Cell Signaling), β-actin (Sigma Aldrich). Rabbit anti-mouse IgG-peroxidase antibody (Sigma Aldrich) and goat anti-rabbit IgG-peroxidase antibody (Sigma Aldrich) were used as the secondary antibody and protein-antibody complexes were visualized by enhanced chemiluminescence system. The Western blotting signals were quantified with ImageJ software (rsbweb.nih.gov/ij/) 14 (link), 15 (link).
Phosph p70s6k
Manufactured by Cell Signaling Technology
Sourced in United States
Phosph-p70s6K is a phospho-specific antibody that detects phosphorylation of p70 S6 kinase at Thr389. It is used to monitor the activation of the mTOR signaling pathway.
Automatically generated - may contain errors
Lab products found in correlation
Rabbit anti mouse igg peroxidase antibody, by Merck Group (2 mentions)
Hybond enhanced chemiluminescence nitrocellulose membrane, by Cytiva (2 mentions)
Phosph mtor, by Cell Signaling Technology (2 mentions)
Phosph akt, by Santa Cruz Biotechnology (2 mentions)
Caspase 3, by Cell Signaling Technology (2 mentions)
P70s6k, by Cell Signaling Technology (2 mentions)
β actin, by Merck Group (2 mentions)
Bca protein assay, by Thermo Fisher Scientific (2 mentions)
Protein kinase b akt, by Santa Cruz Biotechnology (2 mentions)
Goat anti rabbit igg peroxidase antibody, by Merck Group (2 mentions)
Hybond, by Cytiva (2 mentions)
2 protocols using phosph p70s6k
1
Protein Quantification and Western Blotting Protocol
2
Western Blot Quantification Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The protein content was determined using bicinchoninic acid (BCA) protein assay (Pierce Biotechnology, Rockford, IL, USA). About 60-80 µg of protein from the lysates were loaded and fractionated on SDS-PAGE, transferred onto Hybond enhanced chemiluminescence nitrocellulose membranes (Amersham, Little Chalfont, UK) and detected with antibodies against P-gp (GeneTex, Inc. Irvine, CA, USA), the mammalian target of rapamycin (mTOR) (Cell Signaling, Danvers, MA, USA), phosph-mTOR (Cell Signaling), protein kinase B (AKT) (Santa Cruz Biotechnology, Inc. Santa Cruz, CA, USA), phosph-AKT (Santa Cruz Biotechnology, Inc.), p70s6K (Cell Signaling), phosph-p70s6K (Cell Signaling), caspase 3 (Cell Signaling), β-actin (Sigma Aldrich). Rabbit anti-mouse IgG-peroxidase antibody (Sigma Aldrich) and goat anti-rabbit IgG-peroxidase antibody (Sigma Aldrich) were used as the secondary antibody. The protein-antibody complexes were visualized using enhanced chemiluminescence system. The Western blotting signals were quantified with ImageJ software (rsbweb.nih.gov/ij/) 20 (link).
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