Human Cystatin B Quantikine ELISA Kit

    
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  • 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.
Control Available
QC74 , Quantikine Immunoassay Control Set 766 for Human Cystatin B - Please Inquire
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.
 Cystatin B/Stefin B [HRP]
Product Datasheets

Certificate of Analysis Lookup

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Note: Certificate of Analysis not available for kit components.

Certificate of Analysis Lookup

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To download a Certificate of Analysis, please enter the catalog and lot numbers in the search box below.

Note: Certificate of Analysis not available for kit components.

Preparation and Storage
  • 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
Related Research Areas
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

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ELISA Controls
Description Application Cat# Citations Images  

Quantikine Immunoassay Control Set 766 for Human Cystatin B

Ctrl QC74
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Supplemental ELISA Products
Description Application Cat# Citations Images  

Quantikine ELISA Wash Buffer 1

ELISA WA126 6  
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