Human Cystatin B Quantikine ELISA Kit

Catalog # Availability Size / Price Qty
DCYB00
Control Products Available
Human Cystatin B ELISA Standard Curve
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Product Details
Procedure
FAQs
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Human Cystatin B Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (17 uL), EDTA Plasma (17 uL), Heparin Plasma (17 uL), Saliva (10 uL), Urine (13 uL), Human Milk (10 uL)
Sensitivity
0.034 ng/mL
Assay Range
0.2 - 10 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine, Human Milk)
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.

Product Summary

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.

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, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine, Human Milk

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 1.38 2.73 5.47 1.42 2.78 5.4
Standard Deviation 0.049 0.082 0.164 0.148 0.191 0.263
CV% 3.6 3 3 10.4 6.9 4.9

Recovery

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.
Human Cystatin B ELISA Linearity

Data Examples

Human Cystatin B ELISA Standard Curve

Product Datasheets

Preparation and Storage

Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Cystatin B

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.

Entrez Gene IDs:
1476 (Human); 13014 (Mouse)
Alternate Names:
CPI-B; CST6cystatin B (liver thiol proteinase inhibitor)10STFBcystatin-B; CSTB; cystatin B (stefin B); Cystatin B; EPM1; Liver thiol proteinase inhibitor; PME; Stefin B; stefin-B
⚠ 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.

Assay Procedure

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.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 100 µL Assay Diluent
  4.   Add 100 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   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.

FAQs

No product specific FAQs exist for this product, however you may

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