Total protein was extracted from cultured cells using radioimmunoprecipitation (RIPA) lysis solution and protein concentration was assessed. Equal amount of proteins were separated by SDS-PAGE, and then transferred onto PVDF membranes. After being blocked by 5% non-fat milk in 0.1%TBST (Tris-buffered saline), the membranes were incubated with primary antibodies overnight at 4 °C followed by incubation with HRP-conjugated secondary antibodies for 1 h. Protein signal was detected using an enhanced chemiluminescence (ECL) kit. GAPDH was used as an internal control to normalize protein levels. The primary antibodies used in this study include Bcl-2 (Abcam, Cambridge, MA, USA), BAX (Abcam), Pro-caspase-3 (Abcam), cleaved-caspase-3 (Abcam), ACAN (Santa Cruz, USA), COL2A (Santa Cruz), MMP13 (Santa Cruz), LC3 (Proteintech, Chicago, IL, USA), Beclin-1 (Proteintech), p-62 (Proteintech) and GAPDH (Santa Cruz).
Col2a
Manufactured by Santa Cruz Biotechnology
Sourced in United States
COL2A is a laboratory product used for the detection and quantification of the COL2A1 gene, which encodes the type II collagen protein. It is a useful tool for researchers studying the structure and function of cartilage and other connective tissues.
Automatically generated - may contain errors
Lab products found in correlation
Cleaved caspase 3, by Abcam (1 mentions)
Bcl 2, by Abcam (1 mentions)
Bcl2 associated x (bax), by Abcam (1 mentions)
Mmp 13, by Santa Cruz Biotechnology (1 mentions)
Pro caspase 3, by Abcam (1 mentions)
Gapdh, by Santa Cruz Biotechnology (1 mentions)
Beclin 1, by Proteintech (1 mentions)
Polyvinylidene fluoride (pvdf), by Bio-Rad (1 mentions)
P ampkα1, by ZenBio (1 mentions)
Gapdh, by ZenBio (1 mentions)
Il 1β, by Proteintech (1 mentions)
Mmp13, by ZenBio (1 mentions)
Ecl western detection kit, by Thermo Fisher Scientific (1 mentions)
Interleukin 6 (il 6), by ZenBio (1 mentions)
Cyclin d1, by ZenBio (1 mentions)
2 protocols using col2a
1
Western Blot Analysis of Cellular Proteins
2
Chondrocyte Protein Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Rat chondrocytes were lysed by RIPA lysis buffer to extract the total protein which was separated by SDS-PAGE gel electrophoresis and transferred to PVDF (polyvinylidene fluoride) membrane (0.2 μm; BioRad). After 1 h blocking with 5% skimmed milk, the membrane was incubated with primary antibody ACAN (Abcam, ab36861), COL2A (Santa Cruz, sc-7763), IL-1β (Proteintech, 66,737-1-lg), IL-6 (Zen Bio, 347023), MMP-13 (Zen Bio, 511755), PTGS2 (Zen Bio, 383967), Bcl-2 (Zen Bio, 381702), CDK1 (Abcam,ab18), Cyclin D1 (Zen Bio, 382442), p-AMPKα1 (Zen Bio, 310044), AMPKα (Zen Bio, 380431), and GAPDH (Zen Bio, 384404) antibodies overnight at 4°C. After washing 3 times with 1×TBST, the membrane was incubated with horseradish peroxidase-labeled secondary antibody (goat anti-rabbit/mouse IgG, 1:5,000) at room temperature for 1 h. Finally, target protein was visualized by the ECL Western Detection Kit (Thermo Fisher Scientific, United States) and quantified by ImageJ software.
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