Ab32114

Manufactured by Abcam

Sourced in United Kingdom

Ab32114 is a lab equipment product. It is a device used for scientific research and experimentation in a laboratory setting.

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Lab products found in correlation

4 protocols using ab32114

Western Blot Analysis of Protein Signaling

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48 hours later, transfected cells were placed in RIPA lysis buffer on the ice to extract proteins. The concentration of proteins was detected by BCA Protein Assay Kit (Beyotime, Shanghai, China). Equal amounts of protein extracts (20 μg/lane) were loaded in 12% SDS-PAGE and transferred onto a PVDF membrane (Millipore, Boston, MA, USA). The membrane was then incubated with 5% non-fat milk for 1 h at room temperature and the indicated primary antibodies (1:1,000; Abcam, Cambridge, UK) overnight at 4°C against AHNAK2 (ab70053), Bcl-2 (ab32124), Bax (ab32503), p-MEK (ab214445), MEK (ab33918), p-ERK (ab73209), ERK (ab53277), p-P90RSK (ab32203) and P90RSK (ab32114), as well as secondary antibody (1:10,000; Abcam, Cambridge, UK) for 1 h at room temperature. Finally, all bands were probed with an enhanced chemiluminescence (ECL). The gray values of protein bands were scanned with QUANTITY ONE software and recorded by a Bio-Rad camera system (Bio-Rad, USA).

Wang D.W., Zheng H.Z., Cha N., Zhang X.J., Zheng M., Chen M.M, & Tian L.X. (2020). Down-Regulation of AHNAK2 Inhibits Cell Proliferation, Migration and Invasion Through Inactivating the MAPK Pathway in Lung Adenocarcinoma. Technology in Cancer Research & Treatment, 19, 1533033820957006.

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Antibody-based Signaling Pathway Analysis

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Antibodies against PCNA (2586), phosphor-MEK1/2 (9154), phosphor-p90RSK (11989) and phosphor-MSK1 (9595) were obtained from Cell Signaling Technology (CST, MA, USA). Antibodies against AdipoR1 (ab126611), p90RSK (ab32114), cyclinD1 (ab134175), CDK4 (ab108357), CDK6 (ab124821), total-ERK1/2 (ab184699), phospho-ERK1/ (ab76299) and total-MEK1/2 (ab178876) were obtained from Abcam (Cambridge, UK). Recombinant human adiponectin (1065-AP) and recombinant human epidermal growth factor (EGF, 236-EG) were obtained from R&D System.

Fu S., Xu H., Gu M., Liu C., Wan X., Chen Y., Chen Q., Zhou J, & Wang Z. (2017). Lack of adiponectin and adiponectin receptor 1 contributes to benign prostatic hyperplasia. Oncotarget, 8(51), 88537-88551.

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Adiponectin Signaling Pathway Analysis

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Antibodies of Cleaved-caspase 3 (9661), caspase 3 (9662), PCNA (2586), phospho-MEK1/2 (9154), phospho-p90RSK (11989), phospho-MSK1 (9595), phospho-p38 MAPK (9211), Bax (2774) and Bcl-2 (2872) were obtained from Cell Signaling Technology (CST, MA, USA). Antibodies of AdipoR1 (ab126611), AdipoR2 (ab189446), p90RSK (ab32114), cyclinD1 (ab134175), total-ERK1/2 (ab184699), phospho-ERK1/ (ab76299), total-MEK1/2 (ab178876), phospho-AMPK (ab133448) and phospho-mTOR (ab109268) were obtained from Abcam (Cambridge, UK).
Recombinant human adiponectin (1065-AP), recombinant mouse adiponectin (5095-AC) and recombinant human epidermal growth factor (236-EG) were obtained from R&D System. MEK1/2 inhibitor U0126 was obtained from CST (9911).

Fu S., Xu H., Gu M., Liu C., Wang Q., Wan X., Chen Y., Chen Q., Peng Y., Cai Z., Zhou J, & Wang Z. (2017). Adiponectin deficiency contributes to the development and progression of benign prostatic hyperplasia in obesity. Scientific Reports, 7, 43771.

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Time-dependent Protein Expression Analysis

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Treated cells were incubated for 0, 12, 24, 36, or 48 h, then the cellular proteins were lysed by RIPA (Invitrogen, 89900) supplemented with a protease and phosphatase inhibitor cocktail (Invitrogen, 78440), and incubated with SDS-PAGE loading buffer (Reducing) (Cwbio, CW0027) at 100°C for 10 min. Antibodies used for western blotting were as follows: anti-RSK1 p90 (phospho T359 + S363) antibody (1:1,000, ab32413, Abcam), anti-RSK1 p90 antibody (1:1,000, ab32114, Abcam), anti-Phospho-p44/42 MAPK (Erk1/2) (1:1,000, 4370S, Cell signaling), anti- p44/42 MAPK (Erk1/2) (1:1,000, 4695S, Cell signaling), anti-cHSP60 (1:2,000, sc-57840, Santa Cruz), anti-GAPDH (1:1,0000, ab181602, Abcam), anti-rabbit IgG-HRP-linked antibody (1:5,000, 7074S, Cell signaling), and anti-mouse IgG-HRP-linked antibody (1:5,000, 7076S, Cell signaling). Blots were imaged on a ChemiDoc MP Imaging System (Bio-Rad).

Yu X., Xu Q., Chen W., Mai Z., Mo L., Su X., Ou J., Lan Y., Zheng H, & Xue Y. (2022). Rhein inhibits Chlamydia trachomatis infection by regulating pathogen-host cell. Frontiers in Public Health, 10, 1002029.

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