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Human CXCL10/IP-10 Quantikine QuicKit ELISA

Bio-Techne includes R&D Systems | Catalog # QK266

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QK266

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Key Product Details

Assay Length

80 minutes

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Serum (25 µL), EDTA Plasma (25 µL), Heparin Plasma (25 µL)

Sensitivity

4.1 pg/mL

Assay Range

15.6-1000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)

Product Summary for Human CXCL10/IP-10 Quantikine QuicKit ELISA

The Quantikine® QuicKit™ Human CXCL10/IP-10 Immunoassay is a one step, 80-minute solid phase ELISA designed to measure human IP-10 levels in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human IP-10 and antibodies raised against the recombinant protein. Results obtained using natural human IP-10 showed linear curves that were parallel to the standard curves obtained using the QuicKit™ standards. These results indicate that this kit can be used to determine relative mass values for natural human IP-10.

Product Specifications

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Reactivity

Human

Specificity

Natural and recombinant human IP-10.

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 - Samples from apparently healthy volunteers were evaluated for the presence of human IP-10 in this assay. No medical histories were available for the donors used in this study.

Sample TypeMean (pg/mL)Range (pg/mL)Standard Deviation
Serum (n=10)85.649.8-25062.0
EDTA plasma (n=10)99.460.4-20551.8
Heparin plasma (n=10)13783.9-28175.0

Cell Culture Supernates:
Human peripheral blood cells (PBMCs) (1 x 106 cells/mL) were cultured in RPMI supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. Cells were left unstimulated or stimulated with 10 μg/mL PHA for 5 days. Aliquots of the cell culture supernates were removed, assayed for levels of human IP-10, and measured 12,555 pg/mL and 6570 pg/mL, respectively.

PBMCs were cultured in the medium as described above. Cells were left unstimulated or stimulated with 1 μg/mL LPS and 40 ng/mL of recombinant human IFN-gamma for 24 hours. Aliquots of the cell culture supernates were removed, assayed for levels of human IP-10, and measured 174 pg/mL and 958 pg/mL, respectively.

THP-1 human acute monocyte leukemia cells were left in RPMI supplemented with 10% fetal bovine serum, 50 μM beta-mercaptoethanol, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. Cells were left unstimulated or stimulated with 1 μg/mL of recombinant human IFN-gamma for 8 hours and then 1.0 μg/mL LPS was added. Cells were incubated for an additional 18 hours. Aliquots of the cell culture supernates were removed, assayed for levels of human IP-10, and measured 1172 pg/mL and 270,600 pg/mL, respectively. 

Precision

Intra-Assay Precision (Precision within an assay) Two samples of known concentration were tested twenty times on one plate to assess intra-assay precision.

Inter-Assay Precision (Precision between assays) Two samples of known concentration were tested in ten separate assays to assess inter-assay precision.

Cell Culture Supernates, EDTA Plasma, Heparin Plasma, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 1 2
n 20 20 10 10
Mean (pg/mL) 96.7 521 95.3 515
Standard Deviation 1.12 10.6 10.8 42.1
CV% 1.2 2.0 11.3 8.2

Recovery for Human CXCL10/IP-10 Quantikine QuicKit ELISA

The recovery of human IP-10 spiked to three levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 119 110-129
EDTA Plasma (n=2) 86 78-95
Heparin Plasma (n=2) 104 92-118
Serum (n=2) 71 67-76

Linearity

Human IP-10 ELISA Linearity Curve

Scientific Data Images for Human CXCL10/IP-10 Quantikine QuicKit ELISA

Human IP-10 ELISA Standard Curve

Human IP-10 ELISA Standard Curve

Human IP-10 QuicKit Spiked Recovery Competitor Comparison

IP-10 is spiked at three known concentrations throughout the range of the assay and run to measure response of the spiked sample matrix. Plasma recovery is 95% compared to 49% for the top competitor. In spike and recovery experiments, natural samples are spiked with the recombinant target analyte of interest to identify interference caused by sample matrices.

Human IP-10 QuicKit Spiked Linearity Competitor Comparison

IP-10 is spiked at high concentration in various matrices and diluted with appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay. The linearity is between 99%-124% compared to 112%-183% for the top competitor.

Preparation and Storage

Shipping

The product is shipped at ambient temperature. 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: CXCL10/IP-10/CRG-2

IP-10 was originally identified as an IFN-gamma-inducible gene in monocytes, fibroblasts and endothelial cells. The mouse homolog of human IP-10, CRG-2, shares approximately 67% amino acid sequence identity with human IP-10. The amino acid sequence of IP-10 identified the protein as a member of the CXC chemokine subfamily.

Alternate Names

CRG-2, CRG2, IP-10

Entrez Gene IDs

3627 (Human); 15945 (Mouse)

Gene Symbol

CXCL10

Additional CXCL10/IP-10/CRG-2 Products

Product Documents for Human CXCL10/IP-10 Quantikine QuicKit ELISA

Certificate of Analysis

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

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human CXCL10/IP-10 Quantikine QuicKit ELISA

For research use only

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