The Achilles tendon tissue at the edge of the healing site was lysed using RIPA lysis buffer (Beyotime). Equal amounts (20 μg) of protein from each group were separated by SDS-PAGE electrophoresis. Separated proteins were transferred on to a PVDF membrane. Then, primary antibodies against COLI (ab239007, 1:1000, Cambridge, MA, USA), COLIII (ab239007, 1:1000), and GAPDH (ab179467, 1:1000) after blocking and HRP-conjugated secondary antibodies (Abcam; ab7356, 1:5000) were used to incubate with the membrane. All the antibodies were obtained from Abcam (MA, USA). ECL kit (Thermo Fisher Scientific) was applied to visualize the protein bands. β-Actin was used for normalization. The densitometry analysis was performed by IPP 6.0 (Image-Pro Plus 6.0).
Ab239007
Manufactured by Abcam
Sourced in United States
Ab239007 is a primary antibody that can be used for various research applications. It is a purified polyclonal antibody produced in rabbits and targets a specific protein or epitope. The core function of this product is to facilitate the detection and analysis of the target molecule in various experimental techniques.
Automatically generated - may contain errors
Lab products found in correlation
Ecl kit, by Thermo Fisher Scientific (4 mentions)
Ab7356, by Abcam (2 mentions)
Ab179467, by Abcam (2 mentions)
Ripa lysis buffer, by Beyotime (2 mentions)
Ab39012, by Abcam (2 mentions)
β actin, by Cell Signaling Technology (2 mentions)
Bicinchoninic acid protein kit, by Thermo Fisher Scientific (1 mentions)
Ab136668, by Abcam (1 mentions)
Ab118973, by Abcam (1 mentions)
Ab109330, by Abcam (1 mentions)
Ab5666, by Abcam (1 mentions)
Ab219620, by Abcam (1 mentions)
Pvdf membrane, by Thermo Fisher Scientific (1 mentions)
Ab41037, by Abcam (1 mentions)
Bovine serum albumin (bsa), by Beyotime (1 mentions)
Beclin 1, by Cell Signaling Technology (1 mentions)
Cleaved poly adp ribose polymerase, by Cell Signaling Technology (1 mentions)
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
Ab81299, by Abcam (1 mentions)
Ab179785, by Abcam (1 mentions)
4 protocols using ab239007
1
Achilles Tendon Extracellular Matrix Analysis
2
Protein Extraction and Western Blot Analysis
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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.
3
IL-1β Modulates Cell Signaling Pathways
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Recombinant mouse interleukin-1 beta (IL-1β) was acquired from Peprotech (Cranbury, NJ, USA), and dissolved in sterile phosphate-buffered saline (PBS) containing 0.1% bovine serum albumin (BSA; Beyotime Biotechnology, Shanghai, China) at a concentration of 10 µg/ml. Primary antibodies against BCL-2 (B-cell lymphoma 2; no. 124; rabbit monoclonal), BAX (BCL-2-associated X; no. D2E11; rabbit monoclonal), Cleaved caspase-3 (no. Asp175; rabbit monoclonal), cleaved poly(ADP-ribose) polymerase (PARP; no. Asp214; rabbit monoclonal), Beclin1 (no. D40C5; rabbit monoclonal), and β-actin (no. D6A8; rabbit monoclonal) were purchased from Cell Signaling Technology (Danvers, MA, USA). Abcam (Cambridge, UK) provided the antibodies against matrix metalloprotease (MMP13; ab39012), a disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS5; ab41037), and collagen II (ab239007). While LC3 antibody (L8918) was purchased from Sigma-Aldrich.
4
Chondrocyte Inflammation and Degradation
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Recombinant rat IL-1β was acquired from Peprotech (RockyHill, NJ, USA). Abcam (Cambridge, MA, USA) provided the antibodies against yH2ax (ab81299), cGAS (ab179785), STING (ab179775), MMP13 (ab39012), ADAMTS5 (ab41037), Collagen II (ab239007), Aggrecan (ab3778), p21 (ab107099) and p16 (ab51243). Moreover, the Cell Signaling Technology (Beverly, MA, USA) provided the antibodies against β-actin (#3700).
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