Aggrecan Degradation Assay Protocol
Corresponding Organization : University of Oxford
Variable analysis
- Aggrecan (6-B-4) (Abcam, Cambridge, MA, USA)
- ARGXX (BC-3) (Abcam)
- FLAG-M2 (MilliporeSigma, Burlington, MA, USA)
- Acetylated α-tubulin (6-11B-1) (MilliporeSigma)
- Early endosome antigen 1 (EEA-1; Abcam)
- LRP-1 β-light chain (Abcam)
- LRP-1 α-heavy chain (8G1) (Abcam)
- A disintegrin and metalloproteinase (ADAM)-17 (Abcam)
- MMP-13 (Santa Cruz Biotechnology, Dallas, TX, USA)
- MMP14 (anti-catalytic domain EP1264Y) (Abcam)
- Mouse monoclonal anti–MT1-MMP hemopexin domain 222-1D8 (a gift from Yoshi Itoh, Kennedy Institute)
- Arl13b (ProteinTech, Rosemont, IL, USA)
- Anti-AGEG (a kind gift from Hideaki Nagase, University of Oxford)
- Purified aggrecan from bovine cartilage (MilliporeSigma)
- Recombinant human C-terminal His-tagged human receptor-associated protein (RAP) (produced in Escherichia coli using a pET3a-based expression vector)
- The domain deletion mutant, ADAMTS-5 (TS5-3)-flag
- Recombinant MMP-13
- Recombinant IL-1β (PeproTech, London, United Kingdom)
- Phospho-JNK1/2 (Thermo Fisher Scientific, Waltham, MA, USA)
- β-actin (Cell Signaling Technology, Danvers, MA, USA)
- Purified aggrecan from bovine cartilage (MilliporeSigma)
- Recombinant human C-terminal His-tagged human receptor-associated protein (RAP) (produced in Escherichia coli using a pET3a-based expression vector)
- Purified aggrecan from bovine cartilage (MilliporeSigma)
- Recombinant human C-terminal His-tagged human receptor-associated protein (RAP) (produced in Escherichia coli using a pET3a-based expression vector)
Annotations
Based on most similar protocols
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
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!