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Product Info Summary
| SKU: | EK0672 |
|---|---|
| Size: | 96 wells/kit, with removable strips. |
| Reactive Species: | Human |
| Application: | ELISA |
| Sample Types: | cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). |
Product info
Product Name
Human Cathepsin D ELISA Kit PicoKine®
SKU/Catalog Number
EK0672
Size
96 wells/kit, with removable strips.
*Question: How many samples can I assay/run in this kit?
Description
Pathways/functions (putative): lysosomal aspartic endopeptidase involved in proteolysis and remodeling; linked to cell death and tumor biology in literature. Assay: human sandwich ELISA; matrices—cell culture supernatants, cell lysates, serum/plasma (heparin/EDTA); sensitivity <10 pg/mL; range 156–10,000 pg/mL; no detectable cross-reactivity (from product page). Can be profiled with endothelial marker VE-cadherin/CDH5 to study vascular injury (putative).
Storage & Handling
Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)
Cite This Product
Human Cathepsin D ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0672)
Clonality of Antibodies
See Datasheet for details
Standard Protein
Expression system for standard: NS0; Immunogen sequence: L21-L412
Capture/Detection Antibodies
The capture antibody is polyclonal antibody from goat and the detection antibody is polyclonal antibody from goat.
Sensitivity
<10 pg/ml
Assay Range
156 pg/ml - 10,000 pg/ml
Standard Dilution Instructions
See datasheet of EK0672 for more details
Cross-reactivity
There is no detectable cross-reactivity with other relevant proteins.
Reactive Species
EK0672 is reactive to CTSD in Human samples
Validated Sample Types
cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).
Application Guarantee
EK0672 is guaranteed for ELISA in Human by Boster Guarantee
See how Boster Bio validate our ELISA kits: ELISA Validation Information
Background of CTSD
Cathepsin D is a protein that in humans is encoded by the CTSD gene. This proteinase is a member of the peptidase C1 family, having a specificity similar to but narrower than that of pepsin A. It is mapped to 11p15.5. The cDNA encodes a 412-amino acid protein with 20 and 44 amino acids in a pre- and prosegment, respectively. Cathepsin D is one of the lysosomal proteinases. It is ubiquitously expressed and is involved in proteolytic degradation, cell invasion, and apoptosis. Mutations in this gene are involved in the pathogenesis of several diseases, including breast cancer and possibly Alzheimer disease and it has been considered as a breast cancer tumor marker.
Kit Components
| Catalog Number | Description | Quantity |
|---|---|---|
| EK0672-CAP | Anti-Human CTSD Pre-coated 96-well strip microplate | 1 |
| EK0672-ST | Human CTSD Standard | 2 vials, 10 ng/tube |
| EK0672-DA | Human CTSD Biotinylated antibody (100x) | 100ul |
| AR1103 | Avidin-Biotin-Peroxidase Complex (100x) | 100ul |
| AR1106-1 | Sample Diluent | 30ml |
| AR1106-2 | Antibody Diluent | 12ml |
| AR1106-3 | Avidin-Biotin-Peroxidase Diluent | 12ml |
| AR1104 | Color Developing Reagent (TMB) | 10ml |
| AR1105 | Stop Solution | 10ml |
| AR1106-5 | Wash Buffer (25x) | 20ml |
| PLA-SEA | Adhesive plate sealers | 4 |
*The kit components are not available for individual purchase.
Materials Required But Not Included In Kit
- Microplate Reader capable of reading absorbance at 450nm.
- Incubator.
- Automated plate washer (optional).
- Pipettes and pipette tips capable of precisely dispensing 0.5 µl through 1 ml volumes of aqueous solutions.
- Multichannel pipettes are recommended for large amount of samples.
- Deionized or distilled water.
- 500ml graduated cylinders.
- Test tubes for dilution.
Data Examples, Quality Control Data & Sample Dilution
Validation Standard Curve O.D. At 450nm
| Concentration (pg/ml) | 0 | 156 | 312 | 625 | 1250 | 2500 | 5000 | 10000 |
| O.D. | 0.009 | 0.083 | 0.143 | 0.259 | 0.504 | 0.832 | 1.434 | 2.26 |
Data Example Images
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Human Cathepsin D PicoKine ELISA Kit standard curve
Recommended Sample Dilution Ratios
According to our internal validation assays using this ELISA kit, to detect CTSD, Dilution ratio of 1:100, concentration in serum and plasma is 125-250 ng/ml..
Some articles we found to cite concentrations of CTSD in samples: 11331669 (Pubmed IDs).
Intra Assay Consistency & Inter Assay Consistency
We measured random samples of Human Cathepsin D ELISA Kit PicoKine® within the same batch/lot to ensure the consistency of the kits' performances. ELISA intra assay consistency is measured using wells from the same plate/assay kit. ELISA inter assay consistency is measured using wells from different plates from the same batch production/lot.
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 16 | 16 | 16 | 24 | 24 | 24 |
| Mean (pg/ml) | 189 | 1630 | 5138 | 181 | 1712 | 5150 |
| Standard deviation | 13.6 | 91.28 | 339.1 | 13.75 | 111.28 | 427.45 |
| CV (%) | 7.2% | 5.6% | 6.6% | 7.6% | 6.5% | 8.3% |
Specific Publications For Human Cathepsin D ELISA Kit PicoKine® (EK0672)
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Customer Reviews
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Customer Q&As
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7 Customer Q&As for Human Cathepsin D ELISA Kit PicoKine®
Question
Q: Are Boster Bio recombinant proteins and antibodies sterile?
Verified Customer
Verified customer
Asked: 2020-07-16
Answer
A: although the vials are bottled using aseptic techniques, heat-treated vials, and sterile stock solutions, they are not considered or guaranteed to be sterile. If sterile material is required for an experiment, the material can be filtered through a 0.2 micron filter designed for use with biological fluids.
Boster Scientific Support
Answered: 2020-07-16
Question
Q: if the enzyme conjugated Cathepsin D antibodies are mixed with the substrate, will that convert the substrate into the enzymatic reaction product? Or the enzyme function is only activated when the antibody is attached with the Cathepsin D antigen?
Verified Customer
Verified customer
Asked: 2019-05-12
Answer
A: The enzyme is always active. Avoid contaminating the substrate with enzyme prior to the incubation otherwise it compromises the assay with false positive signal.
Boster Scientific Support
Answered: 2019-05-12
Question
Q: can you suggest the dilution ratio of serum samples for detection of Cathepsin D in Human serum? I am trying to measure a multiple analytes and it requires 100ul of diluted samples for each well. We have limited sample quantitys so we like to dilute as much as possible.
Verified Customer
Verified customer
Asked: 2018-10-22
Answer
A: unable to understand the physiological or pathological context of your samples we cannot recommend a dilution ratio without performing a pilot test with your samples. Here is how you can perform a pilot study on your own: perform a serial dilution of your samples on the Cathepsin D ELISA kit to make sure you have a linear ascending curve followed by a plateau, which signifies the samples saturating the detection limit of the kit. Then you can pick the dilution ratios from samples in the linear part of the curve as your experimental dilution ratio.
If you are interested in using our ELISA service, you can also send us your sample and we will take care of everything for you. You can check our service details here: bosterbio.com/services/assay-services/elisa-testing-service
Since you mentioned you have limited samples, our cost effective multiplex ELISA service would fit perfectly for your needs, where we can generate dozens of data points using as little as 25ul sample volume. Information on this service is also in the above link.
Boster Scientific Support
Answered: 2018-10-22
Question
Q: how are cell lysates prepared for use in Picokine® ELISA kits?
Verified Customer
Verified customer
Asked: 2018-09-09
Answer
A: in those Picokine® ELISAs where cell or tissue lysate is a validated sample type, sample preparation instructions for lysate are present in the product insert. Components in lysate and lysis buffer can impact immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.
Boster Scientific Support
Answered: 2018-09-09
Question
Q: can I use citrate plasma as samples in Human Cathepsin D Picokine® ELISA Kit (Catalog # EK0672)?
Verified Customer
Verified customer
Asked: 2018-08-31
Answer
A: Chelating agents such as EDTA, Heparin and Citrate can attach metal ions from the functional domain of Cathepsin D causing disruption of its protein structure. Cathepsin D may be denatured as a result and may compromise the assay's measurements. The chilating sites could also be too close to the epitopes a must for detection and block the antigen antibody reaction. We have tested the Cathepsin D ELISA, treating samples with different anticoagulants and decided that heparin or EDTA can be used for treatment of blood/plasma samples. Do not use other anticoagulents when collecting samples.
Boster Scientific Support
Answered: 2018-08-31
Question
Q: is there any online tool I can use to streamline the data analysis for my ELISA results?
Verified Customer
Verified customer
Asked: 2018-02-09
Answer
A: We have a web based ELISA curve fitting (4pl) and data analysis tool. Please give it a try: bosterbio.com/biology-research-tools/elisa-data-analysis-online. You can also consult our article on ELISA data analysis: bosterbio.com/elisa-data-analysis-instructions
Boster Scientific Support
Answered: 2018-02-09
Question
Q: how can I thaw whole blood sample for Cathepsin D ELISA after freezing?
Verified Customer
Verified customer
Asked: 2018-01-09
Answer
A: do not freeze and thaw whole blood. erythrocytes are fragile and, if frozen and thawed, will undergo hemolysis rendering the samples useless. To keep your blood samples to test Cathepsin D for a later time, you should let the blood clot in glass tubes and separate the serum to freeze for later analysis.
Boster Scientific Support
Answered: 2018-01-09
