Total protein was extracted from cell lines or tissues using RIPA lysis buffer (Beyotime, Shanghai, China). Total protein was quantified using the bicinchoninic acid protein kit (Thermo Fisher Scientific). Protein (40 µg per lane) was separated with 10% SDS-PAGE and transferred onto PVDF membranes (Thermo Fisher Scientific). Subsequently, the membranes were blocked with 5% skimmed milk in TBST for 1 h at room temperature. The membranes were incubated overnight at 4°C with primary antibodies against: IL-6 (Abcam; ab239007, 1 : 1000), IL-1β (Abcam; ab118973, 1 : 1000), TNF-α (Abcam; ab219620, 1 : 1000), LC3 (Abcam; ab136668, 1 : 1000), p62 (Abcam; ab109330, 1 : 1000), Tim-1 (Abcam; ab5666, 1 : 1000) and GAPDH (Abcam; ab179467, 1 : 1000). Then, the membranes were incubated with HRP-conjugated secondary antibodies (Abcam; ab7356, 1 : 5000) for 1 h. Protein bands were visualized using the ECL kit (Thermo Fisher Scientific). β-actin was used as the loading control. IPP 6.0 (Image-Pro Plus 6.0) was used for the densitometry analysis.
Ab118973
Manufactured by Abcam
Ab118973 is a primary antibody product from Abcam. It is a recombinant monoclonal antibody targeted against a specific antigen. The antibody is designed for use in various in vitro laboratory applications.
Automatically generated - may contain errors
Lab products found in correlation
Ecl kit, by Thermo Fisher Scientific (4 mentions)
Ab136668, by Abcam (2 mentions)
Ab109330, by Abcam (2 mentions)
Ab219620, by Abcam (2 mentions)
Ab179467, by Abcam (2 mentions)
Pvdf membrane, by Thermo Fisher Scientific (2 mentions)
Bicinchoninic acid protein kit, by Thermo Fisher Scientific (1 mentions)
Ab7356, by Abcam (1 mentions)
Ab239007, by Abcam (1 mentions)
Ab5666, by Abcam (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Ab3778, by Abcam (1 mentions)
Ab6503, by Abcam (1 mentions)
Bicinchoninic acid (bca), by Thermo Fisher Scientific (1 mentions)
Ab16502, by Abcam (1 mentions)
Radioimmunoprecipitation assay (ripa), by Beyotime (1 mentions)
2 protocols using ab118973
1
Protein Extraction and Western Blot Analysis
2
Quantitative Protein Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
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RIPA (Beyotime, Shanghai, China) was applied to isolate total protein. BCA (Thermo Fisher Scientific) was used to quantify the total protein. SDS-PAGE (10%) was used to separate proteins (40 μg per lane), and then proteins were transferred onto PVDF membranes (Thermo Fisher Scientific). The membranes were incubated with primary antibodies against aggrecan (ABCAm; ab3778, 1:1,000), MMP-1 (ABCAm; ab118973, 1:1,000), MMP13 (ABCAm; ab219620, 1:1,000), PRDX3 (ABCAm; ab136668, 1:1,000), Cyclin D1 (ABCAm; ab16502, 1:1,000), STAT3 (ABCAm; ab109330, 1:1,000), p-STAT3 (ABCAm; ab6503, 1:1,000), and β-actin (ABCAm; ab179467, 1:1,000) after blocked with 5% skimmed milk for 1 h. After that, the membranes were incubated with secondary antibodies (HRP-conjugated, ABCAm; 1:5,000) for 1 h at room temperature. Protein bands were visualized using the enhanced chemiluminescent (ECL) kit (Thermo Fisher Scientific). β-actin was used for normalization. Image-Pro Plus 6.0 was applied for densitometric analysis.
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