Human CXCL10/IP-10 Quantikine QuicKit ELISA

Catalog #: QK266 Datasheet / COA / SDS

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Summary Sample Values Precision Recovery Data Examples Product Datasheets Preparation and Storage Background Related Research Areas

Human CXCL10/IP-10 Quantikine QuicKit ELISA Summary

Assay Length

80 minutes

Sample Type & Volume Required Per Well

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

Sensitivity

4.1 pg/mL

Assay Range

15.6 - 1,000 pg/mL (Cell Culture Supernates, Serum, Plasma, EDTA Plasma, Heparin Plasma)

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 Type	Mean (pg/mL)	Range (pg/mL)	Standard Deviation
Serum (n=10)	85.6	49.8-250	62.0
EDTA plasma (n=10)	99.4	60.4-205	51.8
Heparin plasma (n=10)	137	83.9-281	75.0

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.

Product Summary

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.

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, Serum, EDTA Plasma, Heparin Plasma

	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	11.3	8.2

Recovery

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

Scientific Data

N/A

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Human IP-10 ELISA Standard Curve

ELISA

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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.

ELISA

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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.

Product Datasheets

Product Datasheet

COA

SDS

Preparation and Storage

Shipping

The product is shipped at ambient temperature. 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: 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.

Entrez Gene IDs:

3627 (Human); 15945 (Mouse)

Alternate Names:

C7; chemokine (C-X-C motif) ligand 10; CRG2; CRG-2; CXCL10; gIP-10; IFI10; INP10; IP-10; mob-1; SCYB10

Related Research Areas

Assay Procedure

These assays utilize an anti-tag coated microplate. Standards, samples, and controls are added to plate, followed by subsequent addition of an antibody cocktail. Following a 1 hour incubation, plates are washed and substrate is added and incubated for 20 minutes. Stop solution is then added and plates are read using a standard microplate reader.

QuicKit ELISA assay procedure