NCI-N87, OE-19, AGS, MGC-803, and SGC-7901 cells were purchased from the Cell Bank of the Chinese Academy of Sciences in Shanghai. Dulbecco’s modified Eagle medium (DMEM) (HyClone, Logan, UT, USA) was used. Hon (purity ≥98%) was purchased from Chengdu Best Reagent Co., Ltd. (Chengdu, China), dimethyl sulfoxide (DMSO, meeting the standard of the American Chemical Society) from Beijing Solarbio Science & Technology Co., Ltd. (Beijing, China), and cell-counting kit 8 (CCK-8) from Invitrogen (Carlsbad, CA, USA). Additionally, HER2 antibody (Abcam, Cambridge, UK), PI3K (CST, Danvers, MA, USA), AKT (CST), P21 (Abcam), matrix metalloproteinase (MMP)-9 (Abcam), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Abcam) were used.
Her2 antibody
Manufactured by Abcam
Sourced in United States
The HER2 antibody is a laboratory reagent used in research applications. It specifically binds to the HER2 (human epidermal growth factor receptor 2) protein, which is a receptor tyrosine kinase involved in cell signaling and growth. The HER2 antibody can be used to detect and quantify the presence of HER2 in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Cell counting kit 8 cck 8, by Thermo Fisher Scientific (1 mentions)
Glyceraldehyde 3 phosphate dehydrogenase gapdh, by Abcam (1 mentions)
Matrix metalloproteinase mmp 9, by Abcam (1 mentions)
Dimethyl sulfoxide (dmso), by Solarbio (1 mentions)
Her 2 protein, by Abcam (1 mentions)
N hydroxysuccinimide, by Merck Group (1 mentions)
Cell lysis buffer, by Thermo Fisher Scientific (1 mentions)
Bradford method, by Thermo Fisher Scientific (1 mentions)
Dab kit, by Vector Laboratories (1 mentions)
5 protocols using her2 antibody
1
Anticancer Effects of Hon in Gastric Cell Lines
2
Antibody-Protein Conjugation Protocol
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HER-2 antibody (Ab) and HER-2 protein were obtained from Abcam. Na2WO4 powder was purchased from Merck. The glutamic acid powder was obtained from Sigma. 1-ethyl-3- (3-dimethylami-nopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS) solutions were prepared by dissolving their powders in deionized water, which were bought from sigma. Phosphate buffer saline (PBS, pH = 7.4) was prepared by adding KCl (200 mg), NaCl (8 g), Na2HPO4 (1.44 g), KH2PO4 (245 mg), into the deionized water at room temperature. Sulfuric acid and nitric acid solutions were prepared by dilution of a concentrated solution them using deionized water.
3
Extracting and Quantifying Cellular Proteins
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To extract cellular proteins, cells were incubated in cell lysis buffer (Invitrogen Life Technologies, Carlsbad, CA, USA) containing protease inhibitors on ice for 30 minutes. Cell lysates were clarified by centrifugation at 14,000 rpm at 4°C for 20 min. Protein concentrations were determined via the Bradford method (Thermo Fisher Scientific, Rockford, IL, USA). For western blot analysis, 30 μg of protein extract from each sample was electrophoresed on 10% SDS-PAGE gels and transferred to nitrocellulose membranes. The membranes were probed with HER2 antibody (Abcam, Cambridge, MA, USA), and ß-actin antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA) was used as a loading control. Signals were developed using an ECL chemiluminescence substrate kit (Advansta, Menlo Park, CA, USA).
4
TGFβ-Induced HER2 Regulation by CuB
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Immune-precipitation was performed as described by us previously [67 ]. Briefly, 0.5x106 MDA-MB-231 cells were plated in 100 mm petridish and treated with 10ng/ml TGFβ for 1h prior to the treatment with 50nM CuB. After 48h CuB treatment, whole cell lysates were prepared using RIPA buffer and immune-precipitated with HER2 antibody (Abcam, Cambridge, MA). Immune complexes were resolved on SDS-PAGE and immune-blotted for ITGA6.
5
Immunohistochemical Detection of Her-2 Expression in Paraffin-Embedded Tumor Sections
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Tumors were embedded in paraffin for sectioning. For histological examinations, five micron serious sections were stained by H&E. Immunohistochemical staining was carried out as described previously21 . Briefly, sections were de-paraffinized and rehydrated followed by antigen retrieval using hot citrate buffer. Sections were then blocked in 5% normal horse serum and 1% normal goat serum and incubated with the Her-2 antibody (1:400, Abcam, Cambridge, MA, USA) for 2 h. The DAB Kit (Vector, Burlingame, CA, USA) was used to visualize the staining according to the manufacturer’s instructions.
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