Pi3kca

Manufactured by Cell Signaling Technology

Sourced in United States

PI3KCA is a gene that encodes the catalytic subunit of the phosphoinositide 3-kinase (PI3K) enzyme. PI3K is a key regulator of various cellular processes, including cell growth, proliferation, and survival.

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Lab products found in correlation

Ripa buffer, by Thermo Fisher Scientific (2 mentions) Anti eif4e, by Santa Cruz Biotechnology (1 mentions) Anti pdcd4, by Abcam (1 mentions) Anti actin, by Abcam (1 mentions) Cdc25b, by Cell Signaling Technology (1 mentions) Ccnd3, by Cell Signaling Technology (1 mentions) Imagequant software, by GE Healthcare (1 mentions) Irdye 680lt conjugated secondary antibody, by LI COR (1 mentions) Anti eif4a1, by Abcam (1 mentions) Hnrnpa1, by Abcam (1 mentions) Imagescanner 3, by GE Healthcare (1 mentions) Odyssey system, by LI COR (1 mentions) Immobilon fl transfer membrane, by Merck Group (1 mentions) P erk, by BD (1 mentions) Kras2b, by Proteintech (1 mentions) Ralbp1, by Cell Signaling Technology (1 mentions) Rac1 2, by Cell Signaling Technology (1 mentions) Parp1, by Cell Signaling Technology (1 mentions) β tubulin, by Cell Signaling Technology (1 mentions) Gapdh, by Cell Signaling Technology (1 mentions)

3 protocols using pi3kca

Comprehensive Protein Quantification Protocol

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Blots were probed using the following primary antibodies: anti-eIF4E (Santa Cruz Biotechnology, Santa Cruz, CA, USA; sc-9976), anti-eIF4A1 (Abcam, Cambridge, UK; ab31217), anti-eIF4B (Epitomics, Burlingame, CA, USA; 2232-1), anti-PDCD4 (Abcam, ab80590), anti-actin (Abcam, ab6276), CCND3 (Cell Signaling, Beverly, MA, USA; 2936), PI3KCA (Cell Signaling, 4249), CDC25B (Cell Signaling, 9525), NPM1 (Cell Signaling, 3542), GNAS (Abcam, ab83735), RPL27A (Abcam, ab74731), hnRNPA1 (Abcam, ab4791) and RPS25 (GeneTex, Irvine, CA, USA; 101526). Suitable secondary antibodies were used for chemiluminescence detection. Samples were analyzed from at least two independent experiments.
Films were scanned on an ImageScanner III using LabScan software (GE Healthcare) and proteins were quantified using ImageQuant software (GE Healthcare), or for Licor analysis, IRDye 680LT-conjugated secondary antibody (LI-COR Biosciences, Lincoln, NE, USA) was used, followed by scanning on the Odyssey system (LI-COR Biosciences).

Modelska A., Turro E., Russell R., Beaton J., Sbarrato T., Spriggs K., Miller J., Gräf S., Provenzano E., Blows F., Pharoah P., Caldas C, & Le Quesne J. (2015). The malignant phenotype in breast cancer is driven by eIF4A1-mediated changes in the translational landscape. Cell Death & Disease, 6(1), e1603-.

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Comprehensive Protein Expression Analysis

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Lysates were made using TNN lysis buffer (50 mM Tris-Cl, 250 mM NaCl, 5 mM EDTA, 0.5% NP-40 supplemented with protease inhibitor). 60ug of protein was loaded onto SDS-PAGE gels then transferred onto Immobilon-FL Transfer Membranes (Millipore IPFL00010). The antibodies used were KRAS2B (Proteintech), MYC (Cell signalling Technology), HRAS (Abcam), ERK (BD Pharmingen), p-ERK (p42/44), MET, YAP1, XPO1, DDX6, PARP1, RALBP1, PI3KCA, RAC1/2, β-tubulin, GAPDH (Cell signalling Technology) and β-actin (Sigma A5316). Quantification of western blot images were done by using Image J software.

Singh K., Lin J., Lecomte N., Mohan P., Gokce A., Sanghvi V.R., Jiang M., Grbovic-Huezo O., Burčul A., Stark S.G., Romesser P.B., Chang Q., Melchor J.P., Beyer R.K., Duggan M., Fukase Y., Yang G., Ouerfelli O., Viale A., de Stanchina E., Stamford A.W., Meinke P.T., Rätsch G., Leach S.D., Ouyang Z, & Wendel H.G. (2021). Targeting eIF4A Dependent Translation of KRAS Signaling Molecules. Cancer research, 81(8), 2002-2014.

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Protein Extraction and Western Blot Analysis

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Protein was extracted from cultured cells employing radioimmunoprecipitation assay (RIPA) buffer (ThermoFisher Scientific) following the manufacturer’s instructions, 1x protease and phosphatase inhibitor cocktail (ThermoFisher Scientific) was added to the RIPA buffer. Protein from three biological replicates were pooled together and resolved on 10 or 15% sodium dodecyl sulfate (SDS) polyacrylamide gel. Membranes were blocked with Odyssey blocking buffer (LI-COR Biosciences, Lincoln, NE) and probed with specific antibodies for PI3KCA (4249; Cell Signaling Technology), p-c-RAF1 (Ser338) (9427; Cell Signaling Technology), CCND1 (2922, Cell Signaling Technology), pAkt (Ser473) (4060; Cell Signaling Technology) and GAPDH (sc-32233; Santa Cruz Biotechnology).

Kulkarni P., Dasgupta P., Bhat N.S., Hashimoto Y., Saini S., Shahryari V., Yamamura S., Shiina M., Tanaka Y., Dahiya R, & Majid S. (2020). Role of the PI3K/Akt pathway in cadmium induced malignant transformation of normal prostate epithelial cells.. Toxicology and applied pharmacology, 409, 115308.

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