IHC assay was applied to detect the expressions of β-catenin, type II collagen (Col-II), matrix metalloproteinase 13 (MMP13), aggre-can neoepitope (ACAN) in articular cartilage. Sections were incubated in citrate buffer (0.01 M, pH 6.0, Cat. no. C1010; Solarbio, Beijing, China) at 60°C for 4 hr or Pepsinum (Cat. no. ZLI-9013; ZSGB Biotechnology, Beijing, China) at 37°C for 30min as antigen retrieval. After that, sections were incubated with primary antibodies of Col-II (diluted 1:1,000, ab34712; Abcam, Cambridge, UK), MMP13 (diluted 1:200, ab39012; Abcam), aggrecan (diluted 1:200, ab76956; Abcam), ACAN (diluted 1:200, NB100–74350; Novus, Oakville, Canada) and β-catenin (diluted 1:500, ab32572; Abcam) overnight at 4°C. Second goat antimouse/rabbit antibody (diluted 1:1,000) was added for 20 min, and diaminobenzidine solution was used for detecting positive staining while hematoxylin for counterstaining. A total of four randomly selected fields from at least three different tissue sections were selected for quantification of positive staining. The mean optical density defining as the ratio of integrated optical density to the corresponding cavity area and the rate of the positive cells were evaluated using Image-Pro Plus Software (Media Cybernetics, Silver Spring, MD).
Nb100 74350
Manufactured by Novus Biologicals
Sourced in Canada
The NB100-74350 is an ELISA plate washer. It is designed to automate the washing process in enzyme-linked immunosorbent assay (ELISA) experiments.
Automatically generated - may contain errors
Lab products found in correlation
Ab39012, by Abcam (2 mentions)
Ab76956, by Abcam (2 mentions)
Pepsinum, by Solarbio (2 mentions)
Ab34712, by Abcam (2 mentions)
Image pro plus software, by Media Cybernetics (2 mentions)
Ab32572, by Abcam (2 mentions)
Immobilion western chemiluminescence substrate, by Merck Group (1 mentions)
Anti creb, by Abcam (1 mentions)
Anti pcreb, by Merck Group (1 mentions)
A2066, by Abcam (1 mentions)
R 05071 500, by Advansta (1 mentions)
R 05072 500, by Advansta (1 mentions)
3 protocols using nb100 74350
1
Immunohistochemical Analysis of Cartilage Markers
2
Western Blot Analysis of Metabolic Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Samples were separated by SDS-PAGE, transferred to PVDF membrane, and blocked by 5% skimmed milk solution as previously described [40 (link),42 (link),76 (link)]. The following primary antibodies were used overnight in 1% skimmed milk solution: anti-UCP1 (1:750, R&D Systems, MAB6158), anti-pCREB (1:1000, Merck-Millipore, Burlington, MA, USA, 05-667), anti-CREB (1:1000, Abcam, Cambridge, UK, ab31387), anti-PGC1α (1:1000, Santa Cruz Biotechnology, Dallas, TX, USA, H-300), anti-CKMT2 (1:1000, Novus Biologicals, Centennial, CO, USA, NBP2-13841), anti-CKB (1:1000, Novus Biologicals, NBP1-84460), anti-CRYAB (1:1000, Novus Biologicals, NB100-2519), anti-β actin (1:5000, Novus Biologicals, A2066), anti-OXPHOS (1:1000, Abcam, ab110411), anti-Aggrecan (1:1000, Novus Biologicals, NB100-74350), and anti-ID1 (1:1000, Novus Biologicals, JM92-13). HRP-conjugated goat anti-rabbit (1:10,000, Advansta, San Jose, CA, USA, R-05072-500) or anti-mouse (1:5000, Advansta, R-05071-500) IgG were used as secondary antibodies, respectively. Immunoreactive proteins were visualized by Immobilion Western chemiluminescence substrate (Merck-Millipore). Densitometry was carried out by FIJI.
3
Immunohistochemical Analysis of Articular Cartilage
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
IHC assay was applied to detect the expressions of β‐catenin, type II collagen (Col‐II), matrix metalloproteinase 13 (MMP13), aggrecan neoepitope (ACAN) in articular cartilage. Sections were incubated in citrate buffer (0.01 M, pH 6.0, Cat. no. C1010; Solarbio, Beijing, China) at 60°C for 4 hr or Pepsinum (Cat. no. ZLI‐9013; ZSGB Biotechnology, Beijing, China) at 37°C for 30 min as antigen retrieval. After that, sections were incubated with primary antibodies of Col‐II (diluted 1:1,000, ab34712; Abcam, Cambridge, UK), MMP13 (diluted 1:200, ab39012; Abcam), aggrecan (diluted 1:200, ab76956; Abcam), ACAN (diluted 1:200, NB100‐74350; Novus, Oakville, Canada) and β‐catenin (diluted 1:500, ab32572; Abcam) overnight at 4°C. Second goat anti‐mouse/rabbit antibody (diluted 1:1,000) was added for 20 min, and diaminobenzidine solution was used for detecting positive staining while hematoxylin for counterstaining. A total of four randomly selected fields from at least three different tissue sections were selected for quantification of positive staining. The mean optical density defining as the ratio of integrated optical density to the corresponding cavity area and the rate of the positive cells were evaluated using Image‐Pro Plus Software (Media Cybernetics, Silver Spring, MD).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!