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Human Cystatin C Quantikine ELISA Kit

R&D Systems | Catalog # DSCTC0

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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Serum (50 µL), EDTA Plasma (50 µL), Heparin Plasma (50 µL), Saliva (50 µL), Urine (50 µL), Human Milk (50 µL)

Sensitivity

0.227 ng/mL

Assay Range

3.12-100 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine, Human Milk)
View all Cystatin C ELISA Kits »

Product Summary for Human Cystatin C Quantikine ELISA Kit

The Quantikine Human Cystatin C Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human Cystatin C in cell culture supernates, serum, plasma, saliva, urine, and human milk. It contains NS0-expressed recombinant human Cystatin C and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human Cystatin C showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human Cystatin C.

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 C

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.

Sample Values

Serum/Plasma/Saliva/Urine - Samples from apparently healthy volunteers were evaluated for the presence of human Cystatin C in this assay. No medical histories were available for the donors used in this study.

Sample TypeMean (ng/mL)Range (ng/mL)Standard Deviation (ng/mL)
Serum (n=36)792553-1257161
EDTA plasma (n=36)774560-1173155
Heparin plasma (n=36)786524-1284177
Saliva (n=11)1259103-31841077
Urine (n=12)62.912.6-18843.8

Cell Culture Supernates:
Human peripheral blood cells (1 x 106 cells/mL) were cultured in DMEM supplemented with 5% fetal bovine serum, 50 μM beta -mercaptoethanol, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. Cells were cultured unstimulated or stimulated with 10 μg/mL PHA. Aliquots of the cell culture supernate were removed and assayed for levels of human Cystatin C.

ConditionDay 1 (ng/mL)Day 5 (ng/mL)
UnstimulatedNDND
StimulatedND16
ND=Non-detectable

IMR-90 human lung fibroblast cells were cultured in MEM supplemented with 10% fetal bovine serum, 1.5 g/L sodium bicarbonate, 0.1 mM non-essential amino acids and 0.1 mM sodium pyruvate. An aliquot of the cell culture supernate was assayed for human Cystatin C and measured 23.7 ng/mL.

MCF-7 human breast cancer cells were grown in DMEM/F12 media supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. An aliquot of the cell culture supernate was assayed for human Cystatin C and measured 47.2 ng/mL.

HepG2 hepatocellular carcinoma cells were grown in DMEM supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. An aliquot of the cell culture supernate was assayed for human Cystatin C and measured 264 ng/mL.

Human Milk - Two human milk samples were assayed for human Cystatin C and measured 1456 ng/mL and 2431 ng/mL, respectively. 




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 40 40 40
Mean (pg/mL) 16.2 29.9 52.6 17.2 30.8 60.9
Standard Deviation 1.07 0.92 2.41 1.21 1.53 3.60
CV% 6.6 3.1 4.6 7.0 5.0 5.9

Recovery for Human Cystatin C Quantikine ELISA Kit

The recovery of Cystatin C spiked to levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 96 88-102

Linearity

To assess the linearity of the assay, samples containing high concentrations of Cystatin C were serially diluted with the Calibrator Diluent to produce samples with values within the dynamic range of the assay.

Human Cystatin C ELISA Linearity

Scientific Data Images for Human Cystatin C Quantikine ELISA Kit

Human Cystatin C ELISA Standard Curve

Human Cystatin C 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 C

Cystatin C is a secreted cysteine protease inhibitor that inhibits Cathepsins B, H, K, L, and S. Cystatin C serum concentration correlates closely to the glomerular filtration rate (GFR); elevated levels are associated with coronary artery and cardiovascular disease risk. Dysregulation of Cystatin C can modulate tumor growth and metastasis. In humans, the L68Q variant forms dimers and oligomers more easily than wild type protein and is the cause for hereditary Cystatin C amyloid angiopathy.

Alternate Names

ARMD11, CST3, Gamma-trace, Neuroendocrine basic polypeptide, Post-gamma-globulin

Entrez Gene IDs

1471 (Human); 13010 (Mouse); 25307 (Rat)

Gene Symbol

CST3

Additional Cystatin C Products

Product Documents for Human Cystatin C 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 C 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.

Citations for Human Cystatin C Quantikine ELISA Kit

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  • Human Cystatin C Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Serum
    Verified Customer | Posted 08/19/2021
    Human Cystatin C Quantikine ELISA Kit DSCTC0
  • Human Cystatin C Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Serum and Urine
    Verified Customer | Posted 08/19/2021
    Human Cystatin C Quantikine ELISA Kit DSCTC0
  • Human Cystatin C Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Cell culture supernatant
    Verified Customer | Posted 05/26/2021
    Human Cystatin C Quantikine ELISA Kit DSCTC0
  • Human Cystatin C Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Cell culture supernatant and Human cell line
    Verified Customer | Posted 05/18/2021
  • Human Cystatin C Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Serum
    Verified Customer | Posted 11/27/2018
  • Human Cystatin C Quantikine ELISA Kit
    Name: Jessica Fernando
    Sample Tested: Serum and Urine
    Verified Customer | Posted 12/04/2017

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Protocols

View specific protocols for Human Cystatin C Quantikine ELISA Kit (DSCTC0):

Refer to the product for complete assay procedure.

The conjugate must remain at 2-8 °C during use. Bring all other 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
  3.   Add 100 µL of Assay Diluent to each well.
    4. 50 µL Standard, Control, or Sample
  4.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at 2-8 °C for 3 hours.
  5.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
    6. 200 µL cold Conjugate
  6.   Add 200 µL of cold Conjugate to each well. Cover with a new plate sealer, and incubate at 2-8 °C for 1 hour.
  7.   Aspirate and wash 4 times.
    8. 200 µL Substrate Solution
  8.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
    9. 50 µL Stop Solution
  9.   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.

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