Description
Key facts
Detection method:Colorimetric,
Sample types:Plasma, Tissue Extracts, Cell culture supernatant, Serum, Other biological fluids, Cell Lysate,
Reacts with:Human,
Assay type:Sandwich (quantitative),
Sensitivity:< 5 pg/mL,
Range:156 - 10000 pg/mL,
Assay Platform:Microplate
Product details:
Human Cathepsin B ELISA Kit ab119584 is a sandwich ELISA to measure Human Cathepsin B in serum, plasma, cell culture supernatant, cell lysate, tissue extracts, other biological fluids with a sensitivity of 5 pg/ml.
How the assay works
A Cathepsin B specific goat polyclonal antibody has been precoated onto 96-well plates. Standards and test samples are added to the wells and incubated. A biotinylated detection polyclonal antibody from goat, specific for Cathepsin B is then added followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex is added and unbound conjugates are washed away with PBS or TBS buffer. TMB is then used to visualize the HRP enzymatic reaction. TMB is catalyzed by HRP to produce a blue color product that changes into yellow after adding acidic stop solution. The density of yellow coloration is directly proportional to the Human Cathepsin B amount of sample captured in plate.
Assay Specificity
Our ELISA kits are rigorously validated to ensure the highest level of consistency and reproducibility. Please check the protocol booklet for more details
Human Cathepsin B ELISA Kit ab119584 protocol summary
1. Add standard or sample to each well used. Incubate 37C
2. Add prepared biotin antibody to each well. Incubate at 37C
3. Add prepared Avidin-BiotinPeroxidase Complex (ABC). Incubate at 37C
4. Add TMB to each well. Incubate at room temperature.
5. Add Stop Solution to each well. Read at 450 nm
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
Cathepsin B also known as CTSB or cathepsin beta is a lysosomal cysteine protease belonging to the papain family. This enzyme possesses a molecular weight of approximately 25 to 30 kDa. You can find cathepsin B expressed in various tissues with high levels observed in the liver kidney and spleen. It plays a mechanical role in protein degradation within lysosomes operating optimally in acidic environments. The enzyme has active and inactive forms depending on structural modifications and environmental conditions.
Biological function summary
Cathepsin B participates as a proteolytic enzyme involved in protein turnover and processing. It does not function in isolation but often associates with various other proteases forming complexes essential for efficient substrate breakdown. Cathepsin B assists in the activation of other enzymes by cleaving propeptides therefore facilitating the initiation of cascade reactions. This role is critical in maintaining cellular homeostasis and modulating physiological processes including cell migration and immune response modulation.
Pathways
Cathepsin B significantly contributes to the lysosomal protein degradation pathway and the autophagy-lysosome pathway. It interacts with proteins such as B protein which are important for cellular recycling and energy homeostasis. Its activity supports the MHC class II antigen processing pathway playing an essential part in immune system functionality. Cathepsin B also exhibits interactions with Ctsb protein inhibitors which regulate its proteolytic activity within these pathways.
Cathepsin B has strong associations with cancer progression and neurological disorders such as Alzheimer's disease. Its overexpression and dysregulation are linked to tumor metastasis where it degrades extracellular matrix components promoting invasion. Ctsb protein interactions with corey mazo proteins influence tumor microenvironment and immune evasion. In Alzheimer's disease cathepsin B can influence amyloid beta peptide processing connected to neurodegenerative pathology progression. These roles highlight the potential for cathepsin B as a therapeutic target in addressing these conditions.
