Cystatin B, also known as Stefin B or liver thiol proteinase inhibitor, is a member of family 1 of the cystatin superfamily. It is a 98 amino acid, intracellular inhibitor regulating the activities of cysteine proteases of the papain family such as Cathepsins B, H and L.
Human Cystatin B Quantikine ELISA Kit
R&D Systems | Catalog # DCYB00
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Key Product Details
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 µL), Serum (17 µL), EDTA Plasma (17 µL), Heparin Plasma (17 µL), Saliva (10 µL), Urine (13 µL), Human Milk (10 µL)
Sensitivity
0.034 ng/mL
Assay Range
0.156-10 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine, Human Milk)
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Product Summary for Human Cystatin B Quantikine ELISA Kit
The Quantikine Human Cystatin B Immunoassay is a 4.5 hour solid phase ELISA designed to measure Cystatin B levels in cell culture supernates, serum, plasma, saliva, urine, and human milk. It contains E. coli-expressed recombinant human Cystatin B and antibodies raised against the recombinant protein. Results obtained for naturally occurring human Cystatin B and recombinant human Cystatin B showed linear curves that were parallel to the standard curves obtained using the Quantikine Human Cystatin B Immunoassay standards. These results indicate that this kit can be used to determine relative mass values for natural human Cystatin B.
Product Specifications
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Species
Human
Specificity
Natural and recombinant human Cystatin B
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates, EDTA Plasma, Heparin Plasma, Human Milk, Saliva, Serum, Urine
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 1.38 | 2.73 | 5.47 | 1.42 | 2.78 | 5.40 |
| Standard Deviation | 0.049 | 0.082 | 0.164 | 0.148 | 0.191 | 0.263 |
| CV% | 3.6 | 3.0 | 3.0 | 10.4 | 6.9 | 4.9 |
Recovery for Human Cystatin B Quantikine ELISA Kit
The recovery of Cystatin B spiked to three different levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=4) | 102 | 90-115 |
| EDTA Plasma (n=4) | 96 | 86-103 |
| Heparin Plasma (n=4) | 90 | 81-97 |
| Serum (n=4) | 93 | 83-103 |
Linearity
To assess the linearity of the assay, samples containing high concentrations of Cystatin B were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Scientific Data Images for Human Cystatin B Quantikine ELISA Kit
Human Cystatin B ELISA Standard Curve
Preparation and Storage
Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Cystatin B
Additional Cystatin B Products
Product Documents for Human Cystatin B Quantikine ELISA Kit
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human Cystatin B Quantikine ELISA Kit
For research use only
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.Related Research Areas
Citations for Human Cystatin B Quantikine ELISA Kit
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Sample Tested: Serum and Overexpression cell lysateVerified Customer | Posted 04/20/2022Recombinant human CSTB delivered into mouse using virus vector and assayed from mouse Serum. The mouse serum had low values(both in WT and CSTB KO mice) so signal likely not due to mouse CSTB. Worked as expected in mouse cell line over expressing CSTB.
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Protocols
View specific protocols for Human Cystatin B Quantikine ELISA Kit (DCYB00):
Refer to the product for complete assay procedure.
Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- Add 100 µL of Assay Diluent to each well.
- Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
- Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
- Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
- Aspirate and wash 4 times.
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
- Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
100 µL Assay Diluent
50 µL Standard, Control, or Sample
200 µL Conjugate
200 µL Substrate Solution
50 µL Stop Solution
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