P85 antibody

Manufactured by Cell Signaling Technology

The P85 antibody is a lab equipment product manufactured by Cell Signaling Technology. It is used to detect the p85 subunit of phosphoinositide 3-kinase (PI3K) in various cellular samples.

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Lab products found in correlation

Pierce bca, by Thermo Fisher Scientific (1 mentions) Bis tris protein gel, by Thermo Fisher Scientific (1 mentions) Phospho erk antibody, by Cell Signaling Technology (1 mentions) Pvdf membrane, by Thermo Fisher Scientific (1 mentions) Pierce ecl western blotting substrate, by Thermo Fisher Scientific (1 mentions) Protein g agarose bead, by Thermo Fisher Scientific (1 mentions) Anti v5 tag antibody, by Cell Signaling Technology (1 mentions)

Close spelling variants of this product

Anti-p85 antibody Rabbit polyclonal antibody specific for phosphate-p85 PI3K p85 antibody PI3 kinase p85 antibody Phospho-PI3K p85 (Tyr 458) /p55 (Tyr199) antibody

2 protocols using p85 antibody

Quantification of MAPK Inhibition

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Total protein concentrations were measured by Pierce BCA (ThermoFisher Scientific). Equal amounts of proteins were run on NuPAGE 4 to 12%, Bis-Tris protein gels (ThermoFisher Scientific) and subsequently transferred onto PVDF membranes (ThermoFisher Scientific) by wet electroblotting. Transfer efficiency was checked with Ponceau staining. MAPK inhibition was evaluated via a phospho-ERK antibody (Cell Signaling Technology, #4292, Danvers, MA, USA). Equal protein loading was controlled with the p85 antibody (Cell Signaling Technology, #4370). Antibodies were diluted 1:1000 in 5% BSA in Tris-buffered saline with 0.1% Tween20, incubated overnight at 4 °C, and then probed for one hour at room temperature with a 1:3000 dilution of anti-rabbit (Cell Signaling Technology, #7074) IgG HRP-linked antibody and developed using Pierce ECL Western Blotting Substrate (ThermoFisher Scientific).

Thornton C.E., Hao J., Tamarapu P.P, & Landa I. (2022). Multiple ETS Factors Participate in the Transcriptional Control of TERT Mutant Promoter in Thyroid Cancers. Cancers, 14(2), 357.

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V5-Tag Immunoprecipitation and Western Blot Analysis

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Cell lysates were harvested using ice cold lysis buffer (1% Triton X-100, 10% Glycerol, 100 mM NaCL, 50 mM HEPES, 100 mM Na3VO4, 10 mM NaF, 1 Roche Minitab) and rotated at 4°C for 1 h. Lysates were then clarified by spinning at 10,000 xg at 4°C for 15 min. Protein concentrations were measured using BCA standard curves (Pierce). One μg anti V5-Tag antibody (Cell signaling 13202) was added to 1 mg of clarified cell lysate and rotated at 4°C overnight. 50 μL of Protein G agarose beads (Life technologies 10003D) were added to the lysates and rotated at 4°C for 4 h. Tubes were placed on magnets, washed three times with lysis buffer, and eluted in 20 μL of LDS sample buffer. Lysates were subjected to immunoblot analysis with a p85 antibody (Cell signaling 4257). Unbound lysate was analyzed via western blot for loading controls. Immunoblots were analyzed using ImageJ software.

Croessmann S., Sheehan J.H., Lee K.M., Sliwoski G., He J., Nagy R., Riddle D., Mayer I., Balko J.M., Lanman R., Miller V.A., Cantley L.C., Meiler J, & Arteaga C.L. (2017). PIK3CA C2 domain deletions hyperactivate phosphoinositide 3-kinase (PI3K), generate oncogene dependence and are exquisitely sensitive to PI3Kα inhibitors. Clinical cancer research : an official journal of the American Association for Cancer Research, 24(6), 1426-1435.

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