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Product Info Summary
| SKU: | EK1964 |
|---|---|
| Size: | 96 wells/kit, with removable strips. |
| Reactive Species: | Human |
| Application: | ELISA |
| Sample Types: | cell culture supernatants, serum and plasma (heparin, EDTA) , saliva and urine. |
Product info
Product Name
Human Cystatin D/CST5 ELISA Kit PicoKine®
View all Cystatin D/Cst5 ELISA kits
SKU/Catalog Number
EK1964
Size
96 wells/kit, with removable strips.
*Question: How many samples can I assay/run in this kit?
Description
Human Cystatin D/CST5 ELISA Kit PicoKine™ (96 Tests). Quantitate Human CST5 in cell culture supernatants, serum and plasma (heparin, EDTA) , saliva and urine. Sensitivity: 10pg/ml.
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 Cystatin D/CST5 ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK1964)
Clonality of Antibodies
See Datasheet for details
Immunogen
Expression system for standard: E.coli; Immunogen sequence: G21-V142
Sensitivity
<10 pg/ml
Assay Range
15.6 pg/ml - 1,000 pg/ml
Standard Dilution Instructions
Add 100ul of sample diluent in well #2-#8. Add 200ul standard stock solution to well #1, and serial dilute for well #2-#7 to make a standard curve row. Leave well #8 as blank See datasheet of EK1964 for more details
Cross-reactivity
There is no detectable cross-reactivity with other relevant proteins.
Reactive Species
EK1964 is reactive to CST5 in Human samples
Validated Sample Types
cell culture supernatants, serum and plasma (heparin, EDTA) , saliva and urine.
Application Guarantee
EK1964 is guaranteed for ELISA in Human by Boster Guarantee
See how Boster Bio validate our ELISA kits: ELISA Validation Information
Background of Cystatin D/Cst5
Cystatin-D is a protein that in humans is encoded by the CST5 gene. It is mapped to 20p11.21. The cystatin superfamily encompasses proteins that contain multiple cystatin-like sequences. Some of the members are active cysteine protease inhibitors, while others have lost or perhaps never acquired this inhibitory activity. There are three inhibitory families in the superfamily, including the type 1 cystatins (stefins), type 2 cystatins and the kininogens. The type 2 cystatin proteins are a class of cysteine proteinase inhibitors found in a variety of human fluids and secretions. The cystatin locus on chromosome 20 contains the majority of the type 2 cystatin genes and pseudogenes. This gene is located in the cystatin locus and encodes a protein found in saliva and tears. The encoded protein may play a protective role against proteinases present in the oral cavity.
Kit Components
| Catalog Number | Description | Quantity |
|---|---|---|
| EK1964-CAP | Anti-Human CST5 Pre-coated 96-well strip microplate | 1 |
| EK1964-ST | Human CST5 Standard | 2 vials, 10 ng/tube |
| EK1964-DA | Human CST5 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.
Innovating Scientists Reward
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Data Examples, Quality Control Data & Sample Dilution
Validation Standard Curve O.D. At 450nm
| Concentration (pg/ml) | 0 | 15.6 | 31.2 | 62.5 | 125 | 250 | 500 | 1000 |
| O.D. | 0.013 | 0.063 | 0.113 | 0.208 | 0.361 | 0.713 | 1.211 | 2.038 |
Data Example Images
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Human Cystatin D/CST5 PicoKine ELISA Kitstandard curve
Recommended Sample Dilution Ratios
According to our internal validation assays using this ELISA kit, to detect Cystatin D/Cst5, Dilution ratio of 1:1, concentration in serum and plasma is around 80 pg/ml..
Intra Assay Consistency & Inter Assay Consistency
We measured random samples of Human Cystatin D/CST5 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) | 30 | 215 | 456 | 31 | 287 | 532 |
| Standard deviation | 1.26 | 11.40 | 25.99 | 1.92 | 16.65 | 33.52 |
| CV (%) | 4.2% | 5.3% | 5.7% | 6.2% | 5.8% | 6.3% |
Reproducibility
We ensure reproducibility by testing three samples with differing concentrations of Cystatin D/Cst5 in ELISA kits from four different production batches/lots.
| Lots | Lot 1 (pg/ml) | Lot 2 (pg/ml) | Lot 3 (pg/ml) | Lot 4 (pg/ml) | Mean (pg/ml) | Standard Deviation | CV (%) |
|---|---|---|---|---|---|---|---|
| Sample 1 | 30 | 31 | 27 | 27 | 28 | 1.78 | 6.3% |
| Sample 2 | 215 | 218 | 202 | 231 | 216 | 10.30 | 4.7% |
| Sample 3 | 456 | 461 | 435 | 518 | 467 | 30.74 | 6.5% |
Protein Target Info & Infographic
Gene/Protein Information For CST5 (Source: Uniprot.Org, NCBI)
Gene Name
CST5
Full Name
Cystatin-D
Weight
16.08kDa
Superfamily
cystatin family
Alternative Names
CST5; CSTD; cystatin 5; Cystatin D; Cystatin-5; cystatin-D; cysteine-proteinase inhibitor; MGC71922 CST5 cystatin D cystatin-D|cystatin 5|cysteine-proteinase inhibitor
*if product is indicated to react with multiple species, protein info is based on the gene entry specified above in "species".For more info on CST5, check out the CST5 Infographic
We have 30,000+ of these available, one for each gene! check them out.
In this infographic you will see the following information for CST5: database IDs, super-family, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact us [email protected].
Specific Publications For Human Cystatin D/CST5 ELISA Kit PicoKine® (EK1964)
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No publications found for EK1964
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Customer Reviews
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Customer Q&As
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7 Customer Q&As for Human Cystatin D/CST5 ELISA Kit PicoKine®
Question
Q: if the enzyme conjugated Cystatin 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 Cystatin D antigen?
J. Gandhi
Verified customer
Asked: 2020-09-22
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: 2020-09-22
Question
Q: Can Cystatin D ELISA Kits be used with tissue homogenates (or other non-validated sample types)?
Verified Customer
Verified customer
Asked: 2020-06-11
Answer
A: Unfortunately, Boster Bio has not routinely validated tissue homogenates as a sample type for ELISA kits. This does not mean that ELISA kits are not valid for other sample types than we have tested: it means further investigation is required. One will need to perform a spike and recovery study to determine if an unvalidated sample type will work with a particular kit. To perform a spike and recovery experiment, one should divide a sample into two aliquots. In one of the aliquots, the user should spike in a known amount of the kit standard. a dilution series is performed comparing the spiked versus the unspiked sample. Generally, samples with expected recovery and linearity between 80-120% are considered acceptable. This method can be used to validate any sample type that has not been assessd by Boster Bio. for a more detailed spike and recovery protocol, please contact technical support.
Note: acceptable ranges should be determined individually by each laboratory. Additionally, technical support can help determine if a buffer component is not compatible with a given ELISA kit. please view the Citations tab on the product webpage for peer-reviewed papers utilizing a wide range of sample types. We also have an innovator's reward program where if the user validates our ELISA kits in applications or samples previously not validated by Boster Bio or other users, and share such information with us by submit a review, we will reward the user's efforts with a free antibody or ELISA kit from our catalog. Biocompare.com will also give $20 Amazon giftcard as an additional reward, if the review is submitted there as well.
Boster Scientific Support
Answered: 2020-06-11
Question
A question regarding our previous contact about EK1964: do you have any recommended dilutions for serum and urine samples? I couldn
Verified Customer
Verified customer
Asked: 2020-02-21
Answer
Recommended dilution for serum and urine samples is 1:1.
Boster Scientific Support
Answered: 2020-02-21
Question
Q: can you recommend the dilution ratio of serum samples for detection of Cystatin D in Human plasma ? I am trying to measure a a number of analytes and it requires 100ul of diluted samples for each well. We have low serum volumes so we like to dilute as much as possible.
B. Thompson
Verified customer
Asked: 2019-11-11
Answer
A: without knowing 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 Cystatin 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: 2019-11-11
Question
What is the recommended dilution ratios for serum and urine samples using EK1964?
Verified customer
Asked: 2019-08-01
Answer
The recommended dilution for serum and urine samples using the Human Cystatin D/CST5 ELISA Kit PicoKine™ (EK1964) is 1:1.
Boster Scientific Support
Answered: 2019-08-01
Question
Q: What is the optimal O.D. value for Cystatin D ELISA kit? I used your Cystatin D ELISA on serum samples. For my positive control, I received an O.D. value of 0.826, while my negative control received a value of 0.136. I obtained both of these controls from the ELISA kit, where your kit's typical data shows O.D. values much higher than my positive control and your background is lower. My samples O.D. values are around 0.225 and the highest is only 0.357. can I consider these samples contain Cystatin D even though the O.D. values are not very high?
Verified Customer
Verified customer
Asked: 2019-05-14
Answer
A: The absolute O.D. values may change according to incubation time. The more you incubate the higher the O.D. values are going to be. a point of focus should be is whether your sample O.D. values are statistically significantly higher than your blank values. in your example, you could extend your development time in the substrate incubation step to obtain higher O.D. values, as long as your negative controls' O.D. values are not increasing faster in relation to your positive controls. normally, a sample with O.D. value 2 standard deviations higher than your negative controls can be considered positive. We calculate the sensitivity of this ELISA kit by changeing cutoff O.D. value, calculated as the average of 20 negative controls plus 2 standard deviations of the 20 negative controls, into a concentration. in other words, when we claim this Cystatin D ELISA kit to have sensitivity of 10pg/ml, that means the minimum amount of Cystatin D that can be declared/interpreted as positive by the above standard is 10pg/ml.
Boster Scientific Support
Answered: 2019-05-14
Question
Q: for how much duration can samples (cell cultures, serum, and plasma) be stored and still be stable for measurement Cystatin D using the EK1964 Human Cystatin D Picokine® ELISA Kit?
Verified Customer
Verified customer
Asked: 2019-02-02
Answer
A: Boster Bio does not evaluate sample stability. Variations in sample collection, processing, and storage may change the stabilityof samples. It is recommend to assay sample immediately after collection when possible, or aliquot into single use volumes and store samples frozen. avoid repeat freeze-thaw cycles with the stored samples in order to limit protein degradation.
Boster Scientific Support
Answered: 2019-02-02
