Anti her3

Manufactured by Santa Cruz Biotechnology

Sourced in Denmark

Anti-HER3 is a laboratory reagent used for the detection and analysis of HER3 (Human Epidermal Growth Factor Receptor 3) proteins. It functions as a specific antibody that binds to the HER3 target, enabling researchers to measure and study the expression and distribution of this protein in various biological samples.

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3 protocols using anti her3

Western Blot Analysis of EGFR, HER2, and AKT Signaling

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Cell were lysed in RIPA buffer supplemented with protease and phosphatase inhibitors (Roche). Equal amounts of proteins from cells were separated by 4%–12% SDS-PAGE and were transferred to a PVDF membrane. The blots were blocked for 1 h with 5% Bovine Serum Albumin and then incubated over night with the following primary antibodies: Anti-EGFR (2256, Cell Signaling), Anti-pEGFR (2234, Cell Signaling), anti-HER2 (SC52349, Santa Cruz), anti-pHER2 (2247, Cell Signaling), anti-HER3 (SC81455, Santa Cruz), anti-pHER3 (4791, Cell Signaling), anti-AKT (2920, Cell Signaling), anti-pAKT (4060S, Cell Signaling) anti-P70S6K (49D7, Cell Signaling), anti-pP70S6K (108D2, Cell Signaling), and anti-GAPDH (ab9485, Abcam). Membranes were incubated with their respective secondary antibodies for 1h and analyzed using the enhanced chemiluminescence (ECL) system. Antibody detection and quantification were conducted using the iBright ™ FL1000 (ThermoFisher) and the iBright analysis software.

Sinevici N., Ataeinia B., Zehnder V., Lin K., Grove L., Heidari P, & Mahmood U. (2020). HER3 Differentiates Basal From Claudin Type Triple Negative Breast Cancer and Contributes to Drug and Microenvironmental Induced Resistance. Frontiers in Oncology, 10, 554704.

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Anticancer Compounds Characterization

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Erlotinib was kindly provided by F. Hoffman-La Roche Ltd, gefitinib was provided by AstraZeneca Inc; VS-4718 was provided by Verastem Inc; afatinib, osimertinib, lapatinib, AZD4547, and BIBF1120 were purchased from Selleck Chemicals; SU11274 and picropodophyllin were from Carbiochem; dasatinib was from Bio Vision; SB203580 was from Cayman Chemical; sorafenib was acquired from Toronto Research Chemicals Inc, cisplatin was from Bristol-Myers Squibb Company; and PD173074 was from Sigma-Aldrich.
Anti-HER2 and anti-pHER2 antibodies were purchased from Merck Millipore Corporation, anti-EGFR, anti-pEGFR, anti-pHER3, anti-HER4, anti-pHER4, anti-pc-Met, anti-IGF1Rβ, anti-pIGF1Rβ, anti-PDGFRβ, anti-pPDGFRβ, anti-FGFR1, anti-pFGFR, anti-ERK1/2, anti-pERK1/2, anti-AKT, anti-pAKT, anti-STAT3, anti-pSTAT3, anti-PTEN, anti-SRC, anti-FYN, anti-LYN, anti-YES, anti-LCK, anti-pSRC family (Y416), anti-pSRC (Y527), anti-FAK, anti-pFAK (Y397), anti-pFAK (Y576/577), anti-pFAK (Y925) and anti- EGFR (del E746-A750) antibody were from Cell Signaling Technology, anti-HER3 and anti-c-Met were from Santa Cruz Biotechnology Inc, anti-β-actin was from Abcam, Inc., and anti-α-tubulin was from Sigma-Aldrich.

Murakami Y., Sonoda K., Abe H., Watari K., Kusakabe D., Azuma K., Kawahara A., Akiba J., Oneyama C., Pachter J.A., Sakai K., Nishio K., Kuwano M, & Ono M. (2017). The activation of SRC family kinases and focal adhesion kinase with the loss of the amplified, mutated EGFR gene contributes to the resistance to afatinib, erlotinib and osimertinib in human lung cancer cells. Oncotarget, 8(41), 70736-70751.

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Automated HER2 and HER3 Immunohistochemistry

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Immunohistochemistry was performed using the automated staining system, Ventana NX20 (Roche Diagnostics K.K., Tokyo, Japan) at SRL Co. Ltd. Rabbit polyclonal anti‐HER2 (Dako Denmark A/S, Glostrup, Denmark) and rabbit polyclonal anti‐HER3 (Santa Cruz Biotechnology, Inc., Dallas, TX, USA) antibodies were used. These antibodies were detected using an iVIEW DAB Detection Kit (Roche Diagnostics K.K.). Immunohistochemistry samples without anti‐HER2 or anti‐HER3 antibodies were used as negative controls. Protein expression was evaluated using light microscopy with low (×100), intermediate (×200), and high (×400) magnifications.
Positive expression was defined as strong expression of the examined protein in > 30% of lung cancer cells compared to adjacent lung epithelial cells. Expression was defined as 1+, 2+ and 3+ in cases of weak and membrane staining, complete but moderate membrane staining, and strong and complete staining, respectively.

Kumagai T., Tomita Y., Nakatsuka S., Kimura M., Kunimasa K., Inoue T., Tamiya M., Nishino K., Susaki Y., Kusu T., Tokunaga T., Okami J., Higashiyama M, & Imamura F. (2018). HER3 expression is enhanced during progression of lung adenocarcinoma without EGFR mutation from stage 0 to IA1. Thoracic Cancer, 9(4), 466-471.

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