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.
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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Lab products found in correlation
Anti gapdh antibody, by Abcam (2 mentions)
Enhanced chemiluminescence substrate kit, by Cytiva (1 mentions)
Pvdf membrane, by Beyotime (1 mentions)
Goat anti rabbit igg h l hrp, by Abcam (1 mentions)
Anti mmp13 antibody, by Abcam (1 mentions)
Ripa buffer, by Cell Signaling Technology (1 mentions)
Goat anti mouse igg h l hrp, by Abcam (1 mentions)
Enhanced chemiluminescence, by Thermo Fisher Scientific (1 mentions)
Anti bmp 2 antibody, by Abcam (1 mentions)
Anti smad1 antibody, by Abcam (1 mentions)
Anti col2a1 antibody, by Abcam (1 mentions)
Anti runx2 antibody, by Abcam (1 mentions)
2 protocols using anti acan antibody
1
Protein Expression Analysis of Primary Chondrocytes
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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