Ab190948

Manufactured by Abcam

Sourced in United Kingdom

Ab190948 is a monoclonal antibody that recognizes the EGFR protein. It is suitable for use in various immunoassays.

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Lab products found in correlation

4 protocols using ab190948

Characterization of MCM Protein Expression

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For WB, the detailed procedures were performed as described previously [8] using the following primary antibodies: anti-MCM2 (Abcam, ab4461), anti-MCM3 (Abcam, ab128923), anti-MCM4 (Abcam, ab4459), anti-MCM5 (Abcam, ab75975), anti-MCM6 (Abcam, ab201683), anti-MCM7 (Abcam, ab2360), anti-CDK9 (Abcam, ab76320), anti-CyclinD1 (Abcam, ab16663), anti-CyclinE1 (Abcam, ab33911), anti-p53 (Abcam, ab241566), anti-p21 (Abcam, ab109502), anti-p27 (Abcam, ab32034), and anti-GAPDH (Abcam, ab181602).
For IHC, the standard method was described previously.14 (link) Slides were incubated with primary antibodies (Abs) against MCM2 (Immuway, YM6642), MCM3 (Abcam, ab128923), MCM4 (Immuway, YT2681), MCM5 (Abcam, ab75975), MCM6 (Abcam, ab190948), and MCM7 (Abcam, ab2360).

Niu Y., Shan L., Gao H., Zhang C., Qian Z., Wang Z., Xu X., Zhang X., Wang J., Ma L., Chen L, & Yu Y. (2020). Huaier Suppresses the Hepatocellular Carcinoma Cell Cycle by Regulating Minichromosome Maintenance Proteins. OncoTargets and therapy, 13, 12015-12025.

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Protein Expression Analysis in Lysed Cells

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Cells were lysed with RIPA lysis buffer containing protease inhibitor cocktails (both from Thermo Scientific). Protein concentrations were determined using BCA protein assay kit (Thermo Scientific). Equal amounts of lysates (20-40 µg of protein) were resolved with 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Protein blots were transferred to a nitrocellulose membrane and probed with the corresponding primary antibodies. The membrane was then incubated with appropriate horseradish peroxidase (HRP)-conjugated secondary antibodies, and the protein expression was detected by Immobilon Western Chemiluminescent HRP Substrate (Millipore, Billerica, MA, USA). Antibodies used were all from Abcam (Cambridge, MA, USA): RTKN (Ab154954), CDK1 (Ab18), CDK2 (Ab6538), MCM2 (Ab4461), MCM6 (Ab190948), and GAPDH (Ab8254).

ZHANG W., LIANG Z, & LI J. (2016). Inhibition of rhotekin exhibits antitumor effects in lung cancer cells. Oncology Reports, 35(5), 2529-2534.

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Immunohistochemical Analysis of MCM6 Expression

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The paraffin-embedded tissue sample was divided into 5 μm sections. A section blockade was conducted using the normal goat serum (C-0005, Shanghai Haoran Biological Technology Co., Ltd., Shanghai, China) at room temperature for 20 min and then incubated with the primary rabbit anti-human antibody to MCM6 (ab190948, 1 : 500, Abcam Inc., Cambridge, UK) at 4°C overnight, and with the secondary goat anti-rabbit antibody to Immunoglobulin G (IgG) (ab6785, 1 : 1000, Abcam Inc.) at 37°C for 20 min and with the horseradish peroxidase (HRP)-conjugated streptavidin protein working solution (0343-10000U, Imunbio Co., Ltd., Beijing, China) at 37°C for 20 min. Then, 3’3-diaminobezidin (ST033, Whiga Biotechnology Co., Ltd., Guangzhou, Guangdong, China) was supplemented to the sections for development. Sections were than counter-stained using hematoxylin (PT001, Shanghai Bogoo Biotechnology Co., Ltd., Shanghai, China) for 1 min and immersed in 1% ammonia to develop a blue color gamut, followed by dehydration using alcohol of gradient concentration and xylene clearing. The neutral gum-mounted sections were observed under a microscope (Olympus CX23, Olympus Corp., Japan).

Shao G., Fan X., Zhang P., Liu X., Huang L, & Ji S. (2021). Methylation-dependent MCM6 repression induced by LINC00472 inhibits triple-negative breast cancer metastasis by disturbing the MEK/ERK signaling pathway. Aging (Albany NY), 13(4), 4962-4975.

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Immunohistochemical Analysis of MCM Proteins in Tumor Samples

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Three MCM proteins, namely, MCM2/5/6, were selected for immunohistochemistry (IHC) staining. In brief, pathological sequential slides at 4 μm thickness were sectioned from FFPE tumor blocks and were further deparaffinized and rehydrated. Next, endogenous peroxidase activity was blocked with 3% hydrogen peroxide for 10 min at room temperature. After antigen retrieval, the slides were incubated with primary antibodies and labeled with anti-mouse/rabbit secondary antibody (Dako REAL EnVision Detection System) for 30 min at 37°C, followed by a 3 min incubation in diaminobenzidine solution for protein detection.³¹ (link) The primary antibodies used were anti-MCM2 (ab4461, 1:1600; Abcam, Cambridge, UK), anti-MCM5 (ab17967, 1:1000; Abcam, Cambridge, UK), and anti-MCM6 antibodies (ab190948, 1:800; Abcam, Cambridge, UK).

Zeng L., Liu X.Y., Miao L., Chen K., Xu H., Qin L.J., Li M., Liu K., Feng J, & Wang H.Y. (2023). Risk model based on minichromosome maintenance 2 using objective assessment for predicting survival of neuroblastoma. iScience, 26(2), 105974.

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