Cathepsin D ELISA Kit

Catalog No. ABIN1112571

Product Details Cathepsin D ELISA Kit

Target: Cathepsin D (CTSD)

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 156-10000 pg/mL

Minimum Detection Limit: 156 pg/mL

Application: ELISA

Purpose: For quantitative detection of Cathepsin D in human serum, plasma or cell culture supernatants.

Sample Type: Serum, Plasma, Cell Culture Supernatant

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: 450nm) 3. Precise pipette and disposable pipette tips 4. Automated plate washer 5. ELISA shaker 6. 1.5ml of Eppendorf tubes 7. Plate cover 8. Absorbent filter papers 9. Plastic or glass container with volume of above 1L

Application Details

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)

Target Details for Cathepsin D

Target: Cathepsin D (CTSD)

Alternative Name: Cathepsin D (CTSD Products)

Synonyms: catD ELISA Kit, fb93e11 ELISA Kit, fj17b09 ELISA Kit, wu:fb93e11 ELISA Kit, wu:fj17b09 ELISA Kit, LOC398557 ELISA Kit, Cathd ELISA Kit, CatD ELISA Kit, LOC100136761 ELISA Kit, CTSD ELISA Kit, DKFZp469J0315 ELISA Kit, DDBDRAFT_0205785 ELISA Kit, DDBDRAFT_0215012 ELISA Kit, DDB_0205785 ELISA Kit, DDB_0215012 ELISA Kit, CLN10 ELISA Kit, CPSD ELISA Kit, CD ELISA Kit, cathepsin D ELISA Kit, cathepsin D S homeolog ELISA Kit, lysosomal aspartic protease ELISA Kit, cathepsin D-like ELISA Kit, ctsd ELISA Kit, LOC398557 ELISA Kit, ctsd.S ELISA Kit, CatD ELISA Kit, LOC5567565 ELISA Kit, LOC100136761 ELISA Kit, CTSD ELISA Kit, Cathd ELISA Kit, ctsD ELISA Kit, catd ELISA Kit, Ctsd ELISA Kit, LOC443060 ELISA Kit

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