The chondrocytes were characterized by immunohistochemical staining for type II collagen. The chondrocytes were washed with PBS and fixed with 4% paraformaldehyde for 20 min. After washing thrice with PBS, 0.5% Triton X-100 (Sigma-Aldrich, USA) was added, and the mixture was incubated at 22 ± 3 °C for 15 min. After washing, 3% H2O2 was added, and the resulting mixture was incubated at 22 ± 3 °C for 15 min. The cells were then washed and then incubated with anti-collagen II antibody (1:1000, Proteintech, Chicago, USA) or anti-collagen I antibody (1:1000, NOVUS) overnight. Afterwards, the cells were washed and then treated with horseradish peroxidase (HRP)-labeled secondary antibody (Goat anti-rabbit IgG, Jackson ImmunoResearch Laboratories, Inc.) for 1 h. After washing, the cells were incubated with diaminobenzidine (DAB, Beyotime Biotechnology) for 5 min, and the cells were then redyed with hematoxylin (Sigma-Aldrich). The color of the cells was observed under a microscope at 200× magnification.
Anti collagen 1 antibody
Manufactured by Novus Biologicals
Sourced in United States
The Anti-collagen I antibody is a research tool used to detect and quantify the presence of collagen type I, a major structural protein found in various tissues such as skin, bone, and tendon. This antibody can be utilized in techniques like ELISA, Western Blotting, and Immunohistochemistry to study the expression and distribution of collagen type I in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Diaminobenzidine dab, by Beyotime (1 mentions)
Hematoxylin, by Merck Group (1 mentions)
Anti collagen 2 antibody, by Proteintech (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Goat anti rabbit igg, by Jackson ImmunoResearch (1 mentions)
Losartan, by Merck Group (1 mentions)
Nf κb p65, by Santa Cruz Biotechnology (1 mentions)
Bromodeoxyuridine brdu kit, by Roche (1 mentions)
P akt, by Cell Signaling Technology (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
Anti map2 antibody, by Cell Signaling Technology (1 mentions)
Bca assay, by Beyotime (1 mentions)
β actin, by Absin (1 mentions)
Anti caspase 3 antibody, by Abcam (1 mentions)
Chemiluminescent imaging system, by Bio-Rad (1 mentions)
3 protocols using anti collagen 1 antibody
1
Chondrocyte Characterization by Immunohistochemistry
2
Molecular Mechanisms of Calpain-Mediated Injury
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ANG II, the calpain inhibitor MDL28170, the PI3K inhibitor LY294002, and fluorogenic peptide Suc-Leu-Leu-Val-Tyr-AMC were obtained from Calbiochem (La Jolla, CA). Anti-collagen-I antibody was obtained from Novus Biologicals (Littleton, CO). Antibodies against phospho-Akt (p-Akt) and total-Akt (t-Akt) were purchased from Cell Signaling Technology (Danvers, MA). NF-κB p65 was obtained from Santa Cruz Biotechnology (Dallas, TX). Antibodies against IκB-α and GAPDH were obtained from Epitomics (Burlingame, CA). Anti-calretinin antibody was purchased from BD Transduction (San Jose, CA). Bromodeoxyuridine (BrdU) kit was obtained from Roche (Mannheim, Germany). Enzyme-linked immunosorbent assay (ELISA) kits of ACE were from R&D Systems (Minneapolis, MN). The type 1 ANG II receptor antagonist losartan was obtained from Sigma-Aldrich (St. Louis, MO). Antibody specific for the calpain-mediated cleavage of spectrin (SBDP) was provided by Dr. Kevin K. W. Wang.
3
Spinal Cord Development Protein Analysis
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The embryonic spinal cord tissues at 15 days, 17 days, 19 days, and 21 days after pregnancy in rats were homogenized with RIPA buffer (50 mM Tris-HCl [pH 7.4], 150 mM NaCl, 0.1% SDS, 2 mM EDTA, 1% Na-deoxycholate, and 1% NP-40) supplemented with cocktail protease inhibitor (1:500; Roche, Castle Hill, NSW, Australia). All homogenates were centrifuged at 15,000 RCF for 30 min at 4°C to collect the supernatants, and total protein concentration was assessed by BCA assay (Beyotime Biotechnology). After boiling at 100°C for 5 min, SDS-polyacrylamide gels were used to separate 30 μg protein per well followed by a semidry transfer onto a polyvinylidene difluoride membrane. The membrane was blocked with 5% skimmed milk in TBS for 2 h at room temperature and incubated at 4°C overnight with the following primary antibody solutions: anti-Shh antibody (Novus, rabbit, 1:1,000), anti-MAP2 antibody (Cell Signaling, rabbit, 1:2,000), anti-Collagen I antibody (Novus, rabbit, 1:1,000), and anti-Caspase3 antibody (Abcam, rabbit, 1:500). β-Actin (Absin, rabbit, 1:2,000) was used as a loading control. The membranes were then incubated with the corresponding secondary antibodies (HRP, Goat Anti-Rabbit IgG, 1:5,000) for 1 h at room temperature, and protein bands were visualized and analyzed using a chemiluminescent imaging system (BIO-RAD, USA).
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