NPCs lysates were prepared in RIPA buffer (Thermo) and then extracted protein. Bicinchoninic acid protein assay (Beyotime) was applied for detecting concentrations. Protein was subjected to SDS-PAGE and transferred to a PVDF membrane for immunoblotting analysis. The membranes were incubated with anti-PRKCA (CST, 2056, 1:1000), anti-COL2A1 (Proteintech, 28,459–1-AP, 1:1000) and anti-GAPDH (Proteintech, 60,004–1-Lg, 1:20,000) overnight at 4 °C, and then incubated with goat anti-rabbit IgG-HRP (abcam, ab6721, 1/20000) and goat anti-mouse IgG-HRP (abcam, ab205719, 1:1000) at 24 °C for 2 h. Finally, membranes were exposed to a chemiluminescence imaging analysis and quantified with Image J software.
Anti col2a1
Manufactured by Proteintech
Anti-COL2A1 is a laboratory equipment product from Proteintech. It is a primary antibody that targets the COL2A1 protein, which is a major structural component of cartilage. The core function of this product is to facilitate the detection and analysis of the COL2A1 protein in various research and experimental applications.
Automatically generated - may contain errors
2 protocols using anti col2a1
1
Protein Expression Analysis of NPCs
2
Chondrocyte Transfection and Immunofluorescence
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Chondrocytes were transfected with pEGFP-p67 expression plasmid or incubated with or without 10 μM suramin for 24 hours and then plated on glass coverslips. The cells were washed and fixed with 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.2% Triton X-100, and then blocked with 10% FBS in PBS. The cells were then stained with anti-COL2A1 (1:200) (Proteintech Group) and COL10A1 (Abcam) (1:200) antibody. Images were captured using a fluorescence microscope (NIKON Ti-U).
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