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Anti acan antibody

Manufactured by Abcam

The Anti-ACAN antibody is a laboratory research tool used to detect and study the aggrecan (ACAN) protein. Aggrecan is a key structural component of cartilage and plays a crucial role in the maintenance of the extracellular matrix. The antibody can be used in various immunoassay techniques to identify and quantify ACAN expression in biological samples.

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2 protocols using anti acan antibody

1

Protein Expression Analysis of Primary Chondrocytes

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We used RIPA buffer (Cell Signaling Technology, USA) to extract the total protein of primary chondrocytes. A total of 30 µg protein was loaded into SDS-PAGE gels and subsequently transferred into PVDF membranes (Beyotime). Those membranes were incubated using 5% skim milk (Beyotime) and primary antibodies including anti-COL2A1 antibody (Proteintech), anti-ACAN antibody (Abcam), anti-MMP13 antibody (Abcam), anti-ADAMTS5 antibody (Abcam), and anti-GAPDH antibody (Abcam). Furthermore, these membranes were incubated using secondary antibody (goat antirabbit IgG H&L (HRP) (Abcam) and goat antimouse IgG H&L (HRP) (ab205719, 1:2000, Abcam, UK)) for 1 hour at room temperature. An enhanced chemiluminescence substrate kit (Amersham Pharmacia Biotech, Little Chalfont, UK) was applied to visualize the protein in PVDF membranes.
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2

Protein Expression Analysis in Cartilage Tissue

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The cells were washed with cold PBS 3 times; total protein was extracted with RIPA buffer containing 1% PMSF, and the protein concentration was measured with a quantitative BCA protein kit (Thermo Fisher, USA). The following primary antibodies were used:anti-COL 2A1 antibody (1:1000, Abcam); anti-ACAN antibody (1:1000, Abcam); anti-BMP-2 antibody (1:1000, Abcam); anti-ITG A1 antibody (1:1000, Abcam), anti-Smad1 antibody (1:1000, Abcam), anti-RUNX2 antibody (1:1000, Abcam), and anti-GAPDH antibody (1:1000, Abcam). GAPDH was used for normalization of the data. The membrane was incubated with the corresponding horseradish peroxidase (HRP)-labeled secondary immunoglobulin G conjugate (1:2000, Abcam) at room temperature for 1 h. Protein bands were visualized and detected using an enhanced chemiluminescence (Thermo Fisher, USA) system. The experiments were performed in triplicate.
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