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Phospho erbb3

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Phospho-ERBB3 is a laboratory reagent used to detect and quantify the phosphorylated form of the ERBB3 protein. ERBB3 is a member of the epidermal growth factor receptor (EGFR) family and plays a role in signal transduction pathways. The phospho-ERBB3 product allows researchers to study the activation and regulation of this protein in various biological systems.

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Lab products found in correlation

Phospho erbb2, by Cell Signaling Technology (5 mentions) Phospho akt, by Cell Signaling Technology (5 mentions) Erk1 2, by Cell Signaling Technology (4 mentions) Phospho erk1 2, by Cell Signaling Technology (4 mentions) Erbb3, by Cell Signaling Technology (4 mentions) Phospho egfr, by Cell Signaling Technology (3 mentions) Rabbit anti e cadherin, by Cell Signaling Technology (2 mentions) Goat anti actin, by Santa Cruz Biotechnology (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Erbb3, by Thermo Fisher Scientific (2 mentions) Phospho frs2 α, by Cell Signaling Technology (2 mentions) Phospho fgf receptor, by Cell Signaling Technology (2 mentions) Fgf receptor 3, by Cell Signaling Technology (2 mentions) Erbb2, by Cell Signaling Technology (2 mentions) Pan akt, by Cell Signaling Technology (1 mentions) S6 ribosomal protein, by Cell Signaling Technology (1 mentions) Cell recovery solution, by Corning (1 mentions) Nupage bis tris gel, by Thermo Fisher Scientific (1 mentions) Pan erk1 2, by Cell Signaling Technology (1 mentions) Sc 1616, by Santa Cruz Biotechnology (1 mentions)

Close spelling variants of this product

ErbB3 (29 citations)

9 protocols using phospho erbb3

1

Analyzing Cell Signaling Pathways

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Cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (GIBCO) and incubated at 37 degree in a 5% CO2 humidified incubator. Rabbit anti-E-cadherin, Tubulin, phospho-ERBB2, and phospho-ERBB3 antibodies were from Cell Signaling; rabbit anti-GPC5 (detects N-terminal AAs 46–61: RGLPDSPRAGPDLQVC) was from Biomatic; and goat anti-Actin was from Santa Cruz. All chemicals were from Sigma unless specifically indicated.
Guo L., Wang J., Zhang T, & Yang Y. (2016). Glypican-5 is a tumor suppressor in non-small cell lung cancer cells. Biochemistry and Biophysics Reports, 6, 108-112.
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2

Western Blot Analysis of Receptor Signaling

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Following treatment, cells were lysed in RIPA. Equal amounts of protein were analyzed by SDS-PAGE. Primary antibodies used were as follows: Phospho-FGF Receptor (Tyr653/654, 55H2, #3476S), FGF Receptor 3 (C51F2, #4574S), Phospho-FRS2-α (Tyr436, #3861S), Phospho-ERBB2 (Y1221/1222, #2243S), Phospho-ERBB3 (Y1289, #4791S), Phospho-EGFR (Y1068, 1H12, #2236S), AKT (#9272S), Phospho-AKT (Ser473, D9E, #4060S), Phospho-ERK1/2(T202/Y204, #4370S), ERK1/2 (137F5, #4695S) were from Cell Signaling Technologies. FRS2 (H-91, sc-8318) was purchased from Santa Cruz. ERBB2 (e2-4001) was from NeoMarkers, ERBB3 (#MA5-12675) was from Thermo Scientific. EGFR (#A300-388A) was from Bethyl Antibodies.
Wang J., Mikse O., Liao R.G., Li Y., Tan L., Janne P.A., Gray N.S., Wong K.K, & Hammerman P.S. (2014). Ligand associated ERBB2/3 activation confers acquired resistance to FGFR inhibition in FGFR3 dependent cancer cells. Oncogene, 34(17), 2167-2177.
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3

Protein Extraction and Western Blot Analysis of Organoids

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Organoids were harvested using Cell Recovery Solution (Corning) on ice.
Organoids were lysed using boiling SDS-lysis buffer (50mM Tris-HCL [pH 7.4], 2%
SDS). Protein lysates were separated using 4-12% Bis-Tris NuPage gels (Life
Technologies). Isolation of protein from hN organoids following treatment with
MEK and AKT antagonists required time points of 48 hours or less in order to
obtain sufficient protein quantities. Western blots were probed with the
following antibodies: phospho-ERK1/2 (4370, Cell Signaling), pan-ERK1/2 (4695,
Cell Signaling), phospho-Akt (4060, Cell Signaling), pan-Akt (4685, Cell
Signaling), phospho-ribosomal S6 (4858, Cell Signaling), S6 Ribosomal Protein
(2317, Cell Signaling), pan-EGFR (Abcam, ab2430), phospho-EGFR (3777, Cell
Signaling) ERBB2 (4290, Cell Signaling), phospho-ERBB2 (2247, Cell Signaling),
ERBB3 (12708, Cell Signaling), phospho-ERBB3 (4791, Cell Signaling) Heat Shock
Protein 90 (07-2174, Millipore or 4874, Cell Signaling), actin (sc-1616, Santa
Cruz Biotechnologies) and Kras (sc-30, Santa Cruz Biotechnologies). Loading
control for western blot is Hsp90 unless otherwise indicated.
Ponz-Sarvise M., Corbo V., Tiriac H., Engle D.D., Frese K.K., Oni T.E., Hwang C.I., Öhlund D., Chio I.I., Baker L.A., Filippini D., Wright K., Bapiro T.E., Huang P., Smith P., Yu K.H., Jodrell D.I., Park Y, & Tuveson D.A. (2019). Identification of Resistance Pathways Specific to Malignancy Using Organoid Models of Pancreatic Cancer. Clinical cancer research : an official journal of the American Association for Cancer Research, 25(22), 6742-6755.
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4

Western Blot Analysis of Receptor Signaling

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Following treatment, cells were lysed in RIPA. Equal amounts of protein were analyzed by SDS-PAGE. Primary antibodies used were as follows: Phospho-FGF Receptor (Tyr653/654, 55H2, #3476S), FGF Receptor 3 (C51F2, #4574S), Phospho-FRS2-α (Tyr436, #3861S), Phospho-ERBB2 (Y1221/1222, #2243S), Phospho-ERBB3 (Y1289, #4791S), Phospho-EGFR (Y1068, 1H12, #2236S), AKT (#9272S), Phospho-AKT (Ser473, D9E, #4060S), Phospho-ERK1/2(T202/Y204, #4370S), ERK1/2 (137F5, #4695S) were from Cell Signaling Technologies. FRS2 (H-91, sc-8318) was purchased from Santa Cruz. ERBB2 (e2-4001) was from NeoMarkers, ERBB3 (#MA5-12675) was from Thermo Scientific. EGFR (#A300-388A) was from Bethyl Antibodies.
Wang J., Mikse O., Liao R.G., Li Y., Tan L., Janne P.A., Gray N.S., Wong K.K, & Hammerman P.S. (2014). Ligand associated ERBB2/3 activation confers acquired resistance to FGFR inhibition in FGFR3 dependent cancer cells. Oncogene, 34(17), 2167-2177.
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5

Western Blot Analysis of Signaling Proteins

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Cells were washed twice in cold PBS and lysed with Laemmli sample buffer with β-mercaptoethanol. Proteins were resolved by SDS-PAGE and transferred to PVDF membranes. After blocking in 5% BSA, membranes were incubated with the indicated primary antibodies overnight at 4 °C, followed by incubation with peroxidase-coupled secondary antibodies. Immunoreactivity was detected using horseradish peroxidase (HRP)-conjugated secondary antibodies (CalBioTech, Spring Valley, CA) and chemiluminescence substrate (Thermo Fisher Scientific, Rockford, IL) on a Versadoc imaging system (Bio-Rad). The primary antibodies used were as follows: phospho-ErbB3 (Tyr-1197, #4561), ErbB3 (#4754), PDGFR-β (#3169), AXL (#4939), BRD2 (#5848), BRD4 (#13440), phospho-ERK1/2 (Thr-202/Tyr-204, #9101), ERK1/2 (#9102), and GAPDH (#2118) from Cell Signaling Technology (Danvers, MA); ERK2 (sc-1647) antibody was purchased from Santa Cruz Biotechnology Inc. (Dallas, TX).
Tiago M., Capparelli C., Erkes D.A., Purwin T.J., Heilman S.A., Berger A.C., Davies M.A, & Aplin A.E. (2020). Targeting BRD/BET proteins inhibits adaptive kinome upregulation and enhances the effects of BRAF/MEK inhibitors in melanoma. British Journal of Cancer, 122(6), 789-800.
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6

Comprehensive Cell Culture Immunoblot Protocol

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Cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (GIBCO) and incubated at 37 degree in a 5% CO2 humidified incubator. Rabbit anti-E-cadherin, Tubulin, phospho-ERBB2, and phospho-ERBB3 antibodies were from Cell Signaling; rabbit anti-GPC5 (detects N-terminal AAs 46-61: RGLPDSPRAGPDLQVC) was from Biomatic; and goat anti-Actin was from Santa Cruz. All chemicals were from Sigma unless specifically indicated.
Guo L., Wang J., Zhang T, & Yang Y. (2016). Glypican-5 is a tumor suppressor in non-small cell lung cancer cells. Biochemistry and Biophysics Reports, 6, 108-112.
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7

Antibody-based Immunoblotting Assay

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Antibodies against the following proteins were used, with immunoblotting carried out as previously described [44 (link)]: IRS1 (Upstate Biotechnology, Lake Placid, NY, USA); IRS1 pY612 (Invitrogen), Stat3, Crkl, Jak-1, P85α, CDK16, Cav1, ErbB3, phospho-ErbB3 (Tyr1328; Tyr1325 in mouse), ErbB2, Met, Met pY1234/1235 and pY (P-Tyr-100) (Cell Signaling Technology); phospho-p85 (Tyr467) (GeneTex, Irvine, CA, USA); platelet-derived growth factor receptor α (PDGFRα) and β-actin (Santa Cruz Biotechnology, Santa Cruz, CA, USA); ERBB receptor feedback inhibitor 1 (Errfi1) (Sigma-Aldrich, St Louis, MO, USA).
Ali N.A., Wu J., Hochgräfe F., Chan H., Nair R., Ye S., Zhang L., Lyons R.J., Pinese M., Lee H.C., Armstrong N., Ormandy C.J., Clark S.J., Swarbrick A, & Daly R.J. (2014). Profiling the tyrosine phosphoproteome of different mouse mammary tumour models reveals distinct, model-specific signalling networks and conserved oncogenic pathways. Breast Cancer Research : BCR, 16, 437.
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8

Western Blotting Analysis of Liver Signaling

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Western blotting analysis was performed using 20–100 μg of either nuclear extracts, cytoplasmic extracts, or whole extracts of mouse livers and isolated hepatocytes, as described previously26 (link). The conditioned medium of isolated hepatocytes was also analysed. Antibodies against the following proteins were used: AKT, phospho-AKT (Ser473), CREB, phospho-CREB (Ser133), ERBB1, phospho-ERBB1 (Tyr1068), ERBB3, phospho-ERBB3 (Tyr1289), ERK1/2, phospho-ERK1/2 (Thr202/Tyr204), FoxO1, phospho-FoxO1 (Ser256) (Cell Signaling Technology, Danvers, MA, USA), NRG1α (R&D Systems, Minneapolis, MN, USA), Lamin A/C (Santa Cruz Biotechnology, Santa Cruz, CA, USA), α-Tubulin and Flag (Sigma, St. Louis, MO, USA). Polyclonal antibody against mouse G6Pase and PEPCK was originally established by injecting the corresponding peptides (G6Pase; aa344-357, PEPCK; aa587-600) into guinea pigs (TaKaRa Bio).
Arai T., Ono Y., Arimura Y., Sayama K., Suzuki T., Shinjo S., Kanai M., Abe S.I., Semba K, & Goda N. (2017). Type I neuregulin1α is a novel local mediator to suppress hepatic gluconeogenesis in mice. Scientific Reports, 7, 42959.
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9

Immunoblotting Antibody Panel Analysis

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Immunoblotting experiments were performed using specific antibodies against SREBP-1C (sc-13551; Santa Cruz Biotechnology). The following antibodies were purchased from Cell Signaling Technology: lamin A/C (#2032), phospho-ErbB4 (Tyr1284, #4757), ErbB4 (#4795), phospho-ErbB3 (Tyr1289, #4791), ErbB3 (#12708), phospho-Stat5 (Tyr694, #9359), Stat5 (#94205), phospho-Akt (Ser473; #4060), Akt (#9272), b-actin (#4970), and HSP90 (#4877). Anti-Nrg4 antibody was purchased from Invitrogen (PA5-102641).
Li Y., Jin L., Jiang F., Yan J., Lu Y., Yang Q., Zhang Y., Zhang H., Yu H., Zhang Y., He Z., Zhang R., Yang J, & Hu C. (2021). Mutations of NRG4 Contribute to the Pathogenesis of Nonalcoholic Fatty Liver Disease and Related Metabolic Disorders. Diabetes, 70(10).
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