Ab76523

Manufactured by Abcam

Sourced in United States

Ab76523 is a laboratory reagent produced by Abcam. It is a highly specific antibody that can be used for the detection and quantification of a particular target protein in biological samples. The core function of this product is to serve as a research tool for studying the expression and localization of the target protein.

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Lab products found in correlation

3 protocols using ab76523

Western Blot Analysis of Brain Proteins

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The total protein was extracted with radio-Immunoprecipitation Assay (RIPA) lysate from the brain tissue or A172 cells of each group. The BCA method was used to determine the concentration of the proteins, and 30 µg of the protein samples was separated by sodium dodecyl-sulfate polyacrylamide gel electrophoresis and transferred to the polyvinylidene fluoride (PVDF) membranes using the wet-transfer method. The membranes were blocked with 5% skim milk powder and incubated with primary antibody (1:1,000) at 4 °C overnight. The primary antibody was then washed away, the membranes were invubated with the goat anti-rabbit secondary antibody (Abcam, 1:5,000) at room temperature for 1h. After electrochemical luminescence (ECL) visualization, the relative expression levels of the target bands were derived by the gray scale ratio with the internal reference. The primary antibodies included anti-glial fibrillary acidic protein (GFAP) (Abcam, ab4674), anti-GAPDH (Abcam, ab76523), anti-Bcl-2 (Abcam, ab692), and anti-LC3B (Abcam, ab192890).

Huang N., Gao D., Zhou S, & Huang Z. (2023). Ozone reduces ischemic damage after a stroke by regulating the autophagy of astrocytes. Annals of Translational Medicine, 11(2), 79.

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Bile Protein Analysis in Choledocholithiasis and HCC

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A total of 20 μg of proteins from each patient’s bile (choledocholithiasis, n = 14; HCC, n = 12) were separated by SDS-gel electrophoresis and transferred to PVDF membranes (Millipore, USA). The membranes were then incubated overnight at 4°C with primary antibodies, including anti-PRDX2 (1:1000, Abcam, ab109367), anti-CyclinD1 (1:1000, Abcam, ab16663), anti-cMYC (1:1000, Abcam, ab32072), anti-c-Jun (1:1000, Abcam, ab40766), anti-fra1 (1:1000, Abcam, ab252421), anti-β-catenin (1:1000, Abcam, ab32572), anti-Lamin B1 (1:1000, Abcam, ab16048), anti-GAPDH (1:2000, Abcam, ab76523) and anti-β-actin (1:1000, Abcam, ab8227). Following primary antibody incubation, the membranes were washed and incubated with peroxidase-conjugated secondary antibody (1:2000, Abcam, Cambridge, UK). Subsequently, substrate development was performed using ECL chemiluminescence (Millipore, Bedford, Mass, USA).

YANG X., XIANG X., XU G., ZHOU S., AN T, & HUANG Z. (2023). Silencing of peroxiredoxin 2 suppresses proliferation and Wnt/β-catenin pathway, and induces senescence in hepatocellular carcinoma. Oncology Research, 32(1), 213-226.

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Western Blot Analysis of Protein Targets

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Cells were lysed by RIPA lysis buffer (Beyotime, Shanghai, China). Protein concentration was determined using BCA assay (Beyotime). Proteins were subjected to 10% SDS–polyacrylamide gel electrophoresis. Separated proteins were transferred onto polyvinylidene difluoride (PVDF) membranes (Millipore, Billerica, MA, United States) and immunoblotted with primary antibodies: anti-SOCS2 (ab71676, Abcam), anti-GAPDH (ab76523, Abcam), anti-MMP2 (ab97779, Abcam), and anti-MMP9 (ab38898, Abcam), and secondary antibody: goat anti-rabbit (ab205719; Abcam). Bands were visualized using the EasyBlot ECL Kit (Sangon Biotech, China). The intensity of the blots was quantified by ImageJ (Rawak Software, Germany).

Yang B., Zhao F., Yao L., Zong Z, & Xiao L. (2021). CircRNA circ_0006677 Inhibits the Progression and Glycolysis in Non–Small-Cell Lung Cancer by Sponging miR-578 and Regulating SOCS2 Expression. Frontiers in Pharmacology, 12, 657053.

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