Cathepsin D ELISA Kit

Catalog No. ABIN1112571

The Human Cathepsin D ELISA Kit (ABIN1112571) is a Colorimetric ELISA Kit designed to quantify Human Cathepsin D.

Quick Overview for Cathepsin D ELISA Kit (ABIN1112571)

Target: Cathepsin D (CTSD)

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 156 pg/mL - 10000 pg/mL

Application: ELISA

Sample Type: Serum, Plasma, Cell Culture Supernatant

Product Details Cathepsin D ELISA Kit

Minimum Detection Limit: 156 pg/mL

Purpose: For quantitative detection of Catheps in D in Human serum, plasma,cell culture supernatants.

Analytical Method: Quantitative

Sensitivity: < 10 pg/mL

Components: 1. One 96-well plate pre-coated with anti-human Cathepsin D antibody 2. Lyophilized human Cathepsin D standards: 2 tubes (10ng / tube) 3. Sample / Standard diluent buffer: 30ml 4. Biotin conjugated anti-human Cathepsin D antibody (Concentrated): 130 µl.

Material not included:

1. 37 °C incubator
2. Microplate reader (wavelength: 450 nm)
3. Precise pipette and disposable pipette tips
4. Automated plate washer
5. ELISA shaker
6. 1.5 mL of Eppendorf tubes
7. Plate cover
8. Absorbent filter papers
9. Plastic or glass container with volume of above 1 L

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Comment:

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-Cathepsin D polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-Cathepsin D polyclonal antibody was used as detection antibodies. The standards test samples and biotin conjugated detection antibody were added - the wells subsequently and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were used - visualize HRP enzymatic reaction. TMB was catalyzed by HRP - produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional - the Cathepsin D amount of sample captured in plate. Read the O.D. absorbance at 450 nm in a microplate reader and then the concentration of Cathepsin D can be calculated.

Plate: Pre-coated

Reagent Preparation:

1. Before the experiment, centrifuge each kit component for several minutes to bring down all reagents to the bottom of tubes. 2. It is recommend to measure each standard and sample in duplicate. 3. Do NOT let the plate completely dry at any time! Since the dry condition can inactivate the biological material on the plate. 4. Do not reuse pipette tips and tubes to avoid cross contamination. 5. Do not use the expired components and the components from different batches. 6. To avoid the marginal effect of plate incubation for temperature differences (the marginal wells always get stronger reaction), it is recommend to equilibrate the ABC working solution and TMB substrate for at least 30 min at room temperature (37 °C ) before adding to wells.The TMB substrate (Kit Component 8) is colorless and transparent before use, if not, please contact us for replacement.

Sample Preparation:

Preparation of sample and reagents 1. Sample Isolate the test samples soon after collecting, then, analyze immediately (within 2 hours). Or aliquot and store at -20 °C for long term. Avoid multiple freeze-thaw cycles.
Cell culture supernatants: Centrifuge to remove precipitate, analyze immediately or aliquot and store at -20 °C .
Serum: Coagulate the serum at room temperature (about 4 hours). Centrifuge at approximately 1000 × g for 15 min. Analyze the serum immediately or aliquot and store at -20 °C .
Plasma: Collect plasma with heparin or EDTA as the anticoagulant. Centrifuge for 15 min at 1500 x g within 30 min of collection. Analyze immediately or aliquot and store frozen at -20 °C. Citrate can not be used as anticoagulant here. Note: 1. Coagulate blood samples completely, then, centrifuge, and avoid hemolysis and particle. 2. NaN3 can not be used as test sample preservative, since it is the inhibitor for HRP.

Restrictions: For Research Use only

Handling

Preservative: Sodium azide, Thimerosal (Merthiolate)

Storage: 4 °C,-20 °C

Storage Comment: Store at 2-8°C for 6 months, or at -20°C for 12 months.

Expiry Date: 12 months

Target Details for Cathepsin D

Target: Cathepsin D (CTSD)

Alternative Name: Cathepsin D

Background: Cathepsin D is one of the lysosomal proteinases. It is ubiquitously expressed and is involved in proteolytic degradation, cell invasion, and apoptosis. It is an aspartic protease that depends critically on protonation of its active site Asp residue and gets activated at pH 5 in endosome of hepatocytes where it degrades insulin. Mutations in the CTSD gene are involved in the pathogenesis of several diseases, including breast cancer and possibly Alzheimer disease. It also has been used as a breast cancer tumor marker.

Pathways: Peptide Hormone Metabolism