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Product Info Summary
| SKU: | EK1965 |
|---|---|
| Size: | 96 wells/kit, with removable strips. |
| Reactive Species: | Human |
| Application: | ELISA |
| Sample Types: | cell culture supernatants, serum and plasma (heparin, EDTA). |
Product info
Product Name
Human Cystatin M/Cystatin E/CST6 ELISA Kit PicoKine®
View all Cystatin E/M/CST6 ELISA kits
SKU/Catalog Number
EK1965
Size
96 wells/kit, with removable strips.
*Question: How many samples can I assay/run in this kit?
Description
Human Cystatin M/CST6 ELISA Kit PicoKine® (96 Tests). Quantitate Human CST6 in cell culture supernatants, serum and plasma (heparin, EDTA). Sensitivity: 10pg/ml. The brand Picokine indicates this is a premium quality ELISA kit. Each Picokine kit delivers precise quantification, high sensitivity, and excellent reproducibility. Only our most reliable and effective kits qualify as Picokine, guaranteeing top-tier results for your assays.
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 M/Cystatin E/CST6 ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK1965)
Clonality of Antibodies
See Datasheet for details
Standard Protein
Expression system for standard: E.coli; Immunogen sequence: R29-M149
Sensitivity
<10 pg/ml
Assay Range
46.9 pg/ml - 3,000 pg/ml
Standard Dilution Instructions
See datasheet of EK1965 for more details
Cross-reactivity
There is no detectable cross-reactivity with other relevant proteins.
Reactive Species
EK1965 is reactive to CST6 in Human samples
Validated Sample Types
cell culture supernatants, serum and plasma (heparin, EDTA).
Application Guarantee
EK1965 is guaranteed for ELISA in Human by Boster Guarantee
See how Boster Bio validate our ELISA kits: ELISA Validation Information
Background of Cystatin E/M/CST6
Cystatin-M is a protein that in humans is encoded by the CST6 gene. It is mapped to 11q13.1. 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, where they appear to provide protective functions. This gene encodes a cystatin from the type 2 family, which is down-regulated in metastatic breast tumor cells as compared to primary tumor cells. Loss of expression is likely associated with the progression of a primary tumor to a metastatic phenotype.
Kit Components
| Catalog Number | Description | Quantity |
|---|---|---|
| EK1965-CAP | Anti-Human CST6 Pre-coated 96-well strip microplate | 1 |
| EK1965-ST | Human CST6 Standard | 2 vials, 10 ng/tube |
| EK1965-DA | Human CST6 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 | 46.9 | 93.8 | 187.5 | 375 | 750 | 1500 | 3000 |
| O.D. | 0.020 | 0.058 | 0.098 | 0.192 | 0.392 | 0.775 | 1.429 | 2.010 |
Data Example Images
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Human Cystatin M/CST6 PicoKine ELISA Kitstandard curve
Recommended Sample Dilution Ratios
According to our internal validation assays using this ELISA kit, to detect Cystatin E/M/CST6, Dilution ratio of 1:5, concentration in serum and plasma is around 3 ng/ml..
Intra Assay Consistency & Inter Assay Consistency
We measured random samples of Human Cystatin M/Cystatin E/CST6 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) | 79 | 382 | 1415 | 83 | 409 | 1442 |
| Standard deviation | 4.90 | 17.19 | 80.66 | 4.07 | 23.72 | 90.85 |
| CV (%) | 6.2% | 4.5% | 5.7% | 4.9% | 5.8% | 6.3% |
Reproducibility
We ensure reproducibility by testing three samples with differing concentrations of Cystatin E/M/CST6 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 | 79 | 79 | 75 | 76 | 77 | 1.78 | 2.3% |
| Sample 2 | 382 | 396 | 405 | 372 | 388 | 12.67 | 3.2% |
| Sample 3 | 1415 | 1318 | 1387 | 1330 | 1362 | 39.97 | 2.9% |
Protein Target Info & Infographic
Gene/Protein Information For CST6 (Source: Uniprot.Org, NCBI)
Gene Name
CST6
Full Name
Cystatin-M
Weight
16.511kDa
Superfamily
cystatin family
Alternative Names
Cystatin-M; Cystatin-6; Cystatin-E; CST6 CST6 ECTD15 cystatin E/M cystatin-M|cystatin 6|cystatin M/E|cystatin-E|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 CST6, check out the CST6 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 CST6: 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 M/Cystatin E/CST6 ELISA Kit PicoKine® (EK1965)
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No publications found for EK1965
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Customer Reviews
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Customer Q&As
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8 Customer Q&As for Human Cystatin M/Cystatin E/CST6 ELISA Kit PicoKine®
Question
Q: what is the protocol regarding preparation of cell lysates prepared for use in Picokine® ELISA kits?
Verified Customer
Verified customer
Asked: 2020-11-01
Answer
A: for those Picokine® ELISAs where cell or tissue lysate is a validated sample type, sample preparation instructions for lysate can be found in the product insert. Components in lysate and lysis buffer can change immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.
Boster Scientific Support
Answered: 2020-11-01
Question
Q: if the enzyme conjugated Cystatin E antibodies are mixed with the substrate, will that change the substrate into the enzymatic reaction product? Or the enzyme function is only activated when the antibody is attached with the Cystatin E antigen?
Verified Customer
Verified customer
Asked: 2020-10-10
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: 2020-10-10
Question
Q: What is the optimal O.D. value for Cystatin E ELISA kit? I performed your Cystatin E 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 E even though the O.D. values are not very high?
Verified Customer
Verified customer
Asked: 2020-09-30
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. an important assessment 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 converting 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 E ELISA kit to have sensitivity of 10pg/ml, that means the minimum amount of Cystatin E that can be declared/interpreted as positive by the above standard is 10pg/ml.
Boster Scientific Support
Answered: 2020-09-30
Question
Q: for how much duration can samples (cell cultures, serum, and plasma) be stored and still be stable for measurement Cystatin E using the EK1965 Human Cystatin E Picokine® ELISA Kit?
Verified Customer
Verified customer
Asked: 2020-08-10
Answer
A: Boster Bio does not evaluate sample stability. Variations in sample collection, processing, and storage may affect 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: 2020-08-10
Question
Q: is it okay to use citrate plasma as samples in Human Cystatin E Picokine® ELISA Kit (Catalog # EK1965)?
Verified Customer
Verified customer
Asked: 2020-07-04
Answer
A: Chelating agents such as EDTA, Heparin and Citrate can bind metal ions from the functional domain of Cystatin E causing disruption of its protein structure. Cystatin E may be denatured as a result and may compromise the assay's measurements. The chilating sites could also be too close to the epitopes needed for detection and block the antigen antibody reaction. We have tested the Cystatin E ELISA, treating samples with a number of 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: 2020-07-04
Question
Q: Are Boster Bio recombinant proteins and antibodies sterile?
Verified Customer
Verified customer
Asked: 2019-04-21
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 a must for an experiment, the material can be filtered through a 0.2 micron filter designed for use with biological fluids.
Boster Scientific Support
Answered: 2019-04-21
Question
Q: Can Cystatin E ELISA Kits be used with tissue homogenates (or other non-validated sample types)?
Verified Customer
Verified customer
Asked: 2018-08-10
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 evaluated 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 check 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: 2018-08-10
Question
Q: how to proceed with the analysis of ELISA data? I have performed Cystatin E level in plasma.
Verified Customer
Verified customer
Asked: 2018-03-26
Answer
A: please read this article on ELISA data analysis. bosterbio.com/ELISA-data-analysis-instructions. Boster also provides a free tool that you can use to analyze ELISA data. bosterbio.com/biology-research-tools/ELISA-data-analysis-online
Boster Scientific Support
Answered: 2018-03-26
