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Pi3 kinase p85 19h8

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PI3 Kinase p85 (19H8) is a monoclonal antibody that recognizes the p85 regulatory subunit of phosphoinositide 3-kinase (PI3 kinase). PI3 kinase is an important signaling enzyme that plays a role in various cellular processes.

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Lab products found in correlation

Total mapk, by Cell Signaling Technology (2 mentions) P44 42 mapk erk1 2, by Cell Signaling Technology (2 mentions) Total mek1 2, by Cell Signaling Technology (2 mentions) Phospho mek1 2, by Cell Signaling Technology (2 mentions) Phospho fgf receptor tyr653 654, by Cell Signaling Technology (2 mentions) Fgf receptor 2 d4l2v, by Cell Signaling Technology (2 mentions) Phospho pi3 kinase p85 tyr458 p55 tyr199, by Cell Signaling Technology (2 mentions) Total akt, by Cell Signaling Technology (2 mentions) Phospho akt ser473, by Cell Signaling Technology (2 mentions) β actin, by Cell Signaling Technology (2 mentions) Ab109235, by Abcam (1 mentions) Jak2 d2e12, by Cell Signaling Technology (1 mentions) Phosphatase inhibitor cocktail, by Cell Signaling Technology (1 mentions) Phospho p38 map kinase, by Cell Signaling Technology (1 mentions) P38mapk d13e1, by Cell Signaling Technology (1 mentions) Aprotinin, by Merck Group (1 mentions) Leupeptin, by Merck Group (1 mentions) Ripa lysis buffer, by Beyotime (1 mentions) Pfrs2, by Cell Signaling Technology (1 mentions) Anti frs2, by Abcam (1 mentions)

3 protocols using pi3 kinase p85 19h8

1

Analyzing Protein Signaling Cascades

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The CMECs were homogenized in RIPA lysis buffer (#P0013C; Beyotime) containing 1X Phosphatase Inhibitor Cocktail (#5870S; Cell Signaling Technology, Beverly, MA, USA) and 1 µg/ml each of aprotinin (A1153; Sigma-Aldrich) and leupeptin (#L2884; Sigma-Aldrich). Forty micrograms of protein was separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), transferred to polyvinylidene difluoride (PVDF) membranes, and then probed with antibodies for XO (#ab109235; Abcam), phospho-PI3 kinase p85 (Tyr458)/p55 (Tyr199) (#4228), PI3 kinase p85 (19H8) (#4257), phospho-p38 MAP kinase (Thr180/Thr182) (#4631), p38 MAPK (D13E1) (#8690), JAK2 (D2E12) (#3230) and phospho-JAK2 (Tyr1007/1008) (C80C3) (#3776) (all from Cell Signaling Technology, Beverly, MA, USA). The same membranes were reprobed with an antibody for tubulin (#AT819; Beyotime). The blotting film was quantified using a scanner and a densitometry program (ImageJ; https://imagej.nih.gov/ij/index.html) (24 ). To quantify the phosphor-specific signal in the activated samples, the background was subtracted and the band was normalized to the amount of tubulin or total target protein in the lysate.
Zhang Y.Q., Hu S.Y., Chen Y.D., Guo M.Z, & Wang S. (2016). Hepatocyte growth factor inhibits hypoxia/reoxygenation-induced activation of xanthine oxidase in endothelial cells through the JAK2 signaling pathway. International Journal of Molecular Medicine, 38(4), 1055-1062.
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2

Western Blot Antibody Profiles

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Western blot assays were performed using established protocols with the following antibodies: phospho-Akt (Ser473) 1:1000 (Cell Signaling 9271), Total Akt 1:1000 (Cell Signaling 9272), phospho-MEK1/2 1:5000 (Cell Signaling 9154), Total MEK1/2 1:5000 (Cell Signaling 9122), phospho-FGF Receptor (Tyr653/654) 1:500 (Cell Signaling 3471), FGF Receptor 2 (D4L2V) 1:500 (Cell Signaling 23328), p44/42 MAPK (Erk1/2) 1:5000 (Cell Signaling 9101), Total MAPK 1:5000 (Cell Signaling 9102), pFRS2 1:1000 (Cell Signaling 3864), anti-FRS2 (abcam ab10425), phospho-PI3 Kinase p85 (Tyr458)/p55 (Tyr199) 1:1000 (Cell Signaling 4228), PI3 Kinase p85 (19H8) 1:000 (Cell Signaling 4257), β-actin 1:10000 (Cell Signaling 4967) and GAPDH 1:10000 (Santa Cruz sc-25778).
Krook M.A., Barker H., Chen H.Z., Reeser J.W., Wing M.R., Martin D., Smith A.M., Dao T., Bonneville R., Samorodnitsky E., Miya J., Freud A.G., Monk J.P., Clinton S.K, & Roychowdhury S. (2019). Characterization of a KLK2-FGFR2 fusion gene in two cases of metastatic prostate cancer. Prostate cancer and prostatic diseases, 22(4), 624-632.
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3

Signaling Pathway Protein Analysis

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Western blot assays were carried out using established protocols and probed with the following antibodies: phospho-Akt (Ser473) 1:1000 (Cell Signaling 9271), Total Akt 1:1000 (Cell Signaling 9272), phospho-MEK1/2 1:5000 (Cell Signaling 9154), Total MEK1/2 1:5000 (Cell Signaling 9122), p44/42 MAPK (Erk1/2) 1:5000 (Cell Signaling 9101), Total MAPK 1:5000 (Cell Signaling 9102), phospho-FGF Receptor (Tyr653/654) 1:500 (Cell Signaling 3471), FGF Receptor 2 (D4L2V) 1:500 (Cell Signaling 23328), phospho-PLCγ1 (Tyr783) 1:1000 (Cell Signaling 14008), PLCγ1 (D9H10) 1:1000 (Cell Signaling 5690), phospho-FRS2-α (Tyr196) 1:1000 (Cell Signaling 3864), FRS2 1:1000 (abcam 10425), phospho-PI3 Kinase p85 (Tyr458)/p55 (Tyr199) 1:1000 (Cell Signaling 4228), PI3 Kinase p85 (19H8) 1:000 (Cell Signaling 4257), β-actin 1:10000 (Cell Signaling 4967).
Krook M.A., Bonneville R., Chen H.Z., Reeser J.W., Wing M.R., Martin D.M., Smith A.M., Dao T., Samorodnitsky E., Paruchuri A., Miya J., Baker K.R., Yu L., Timmers C., Dittmar K., Freud A.G., Allenby P, & Roychowdhury S. (2019). Tumor heterogeneity and acquired drug resistance in FGFR2-fusion-positive cholangiocarcinoma through rapid research autopsy. Cold Spring Harbor Molecular Case Studies, 5(4), a004002.
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