Description
Key facts
Detection method:Colorimetric,
Sample types:Plasma, Tissue Extracts, Cell culture supernatant, Serum, Other biological fluids,
Reacts with:Human,
Assay type:Sandwich (quantitative),
Sensitivity:< 10 pg/mL,
Range:156 - 10000 pg/mL,
Assay Platform:Microplate
Product details:
Human Cathepsin D ELISA Kit ab119586 is a sandwich ELISA to measure Human Cathepsin D in serum, plasma, cell culture supernatant, tissue extracts, other biological fluids with a sensitivity of 10 pg/ml.
How the assay works
A Cathepsin D specific mouse monoclonal 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 D 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 D 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 D ELISA Kit ab119586 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 D also known as CTSD is a protein with a mass of approximately 45 kDa. It functions as an aspartyl protease and is expressed in lysosomes across various tissues including the liver and kidneys. This enzyme acts by cleaving peptide bonds within proteins which is essential for protein degradation and turnover. Cathepsin D exists as precursor forms that become activated in the acidic environment of the lysosome. It plays a critical role in normal cellular processes by maintaining protein homeostasis.
Biological function summary
The enzymatic activity of Cathepsin D is important for cellular maintenance and apoptosis. This protease does not act within larger protein complexes but contributes to the degradation of extracellular and intracellular proteins. It mediates processes like antigen processing where it deconstructs proteins into peptides that are presented on major histocompatibility complex (MHC) molecules. ELISA tests can quantify its expression levels sometimes termed as CTSD activity in various biological samples offering insights into its role within cellular environments.
Pathways
Cathepsin D involvement includes the lysosomal degradation pathway and the apoptotic signaling pathway. In the lysosomal degradation pathway Cathepsin D breaks down proteins and peptides a process important for cellular recycling and energy release. It interacts with other lysosomal enzymes such as Cathepsin B in this pathway ensuring comprehensive breakdown of cellular waste. The apoptotic signaling pathway involves the regulation of programmed cell death where Cathepsin D can influence the activation of downstream proteins like Bcl-2 and Bax which control cell survival.
The overexpression of Cathepsin D links to breast cancer and Alzheimer's disease. In breast cancer increased Cathepsin D expression correlates with tumor progression and metastasis influencing tumor behavior through interactions with other proteins involved in cell proliferation. Alzheimer's disease features the involvement of Cathepsin D in the breakdown of amyloid precursor protein which relates to amyloid beta plaque accumulation. The abnormal activity of Cathepsin D in these disorders makes it a potential target for therapeutic antibodies such as CTSD antibodies which aim to regulate its function.
