Human S100A8/S100A9 Heterodimer ELISA Kit - Quantikine Best Seller
R&D Systems, part of Bio-Techne | Catalog # DS8900
Key Product Details
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 µL), Tissue Lysates (50 µL), Serum (10 µL), EDTA Plasma (10 µL), Heparin Plasma (10 µL), Saliva (10 µL), Urine (50 µL), Fecal Extract (50 µL), Human Milk (10 µL)
Sensitivity
0.215 ng/mL
Assay Range
0.625-40 ng/mL (Cell Culture Supernates, Tissue Lysates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine, Human Milk)
Product Summary for Human S100A8/S100A9 Heterodimer Quantikine ELISA Kit
The Quantikine Human S100A8/S100A9 Heterodimer Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human S100A8/S100A9 Heterodimer in cell culture supernates, tissue lysastes, serum, plasma, saliva, urine, human milk, and fecal extract. It contains E. coli-expressed recombinant human S100A8/S100A9 Heterodimer and antibodies raised against the recombinant factor. Results obtained using natural human S100A8/S100A9 Heterodimer 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 S100A8/S100A9 Heterodimer.
Product Specifications
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Reactivity
Human
Specificity
Natural and recombinant human S100A8/S100A9 Heterodimer.
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/Human Milk - Samples from apparently healthy volunteers were
evaluated for the presence of human S100A8/S100A9 Heterodimer in this assay. No medical
histories were available for the donors used in this study.
Cell Culture Supernates:
THP-1 human acute monocytic leukemia cells were cultured in RPMI 1640 supplemented with 10% fetal bovine serum and 2 mM L-glutamine. Cells were cultured unstimulated or stimulated with 200 nM PMA until confluent. An aliquot of the cell culture supernate was removed, assayed for human S100A8/S100A9 Heterodimer, and measured 11.6 ng/mL and 38.0 ng/mL, respectively.
Fecal Extracts - Three fecal samples were extracted by adding 5 mL extraction buffer (0.1 M Tris, 0.015 M NaCl, 1.0 M Urea, 1.0 mM CaCl2, 0.1 M Citric Acid Monohydrate, 5 mg/mL BSA, and 0.25% Gentamycin Sulfate at pH 8.0) into 100 mg sample. Samples were vortexed for 10 minutes and filtered through a 0.2 μm filter. Samples were assayed for human S100A8/S100A9 Heterodimer and measured 81.2 ng/mL, 8.71 ng/mL, and 1175 ng/mL.
| Sample Type | Mean (ng/mL) | Range (ng/mL) | Standard Deviation (ng/mL) |
| Serum (n=72) | 2015 | 481-6540 | 1219 |
| EDTA plasma (n=40) | 473 | 127-1395 | 295 |
| Heparin plasma (n=40) | 830 | 298-1640 | 314 |
| Saliva (n=10) | 7271 | 2137-18,960 | 5336 |
| Urine (n=10) | 102 | 0.858-359 | 99.7 |
| Human milk (n=10) | 1308 | 34.0-5720 | 1876 |
Cell Culture Supernates:
Human peripheral blood leukocytes (PBL; 1x106
) were cultured in RPMI 1640 and
supplemented with 10% fetal bovine serum. Cells were cultured unstimulated or stimulated
with 10 μg/mL PHA for 1 or 5 days. Aliquots of the cell culture supernates were removed and
assayed for levels of human S100A8/S100A9 Heterodimer.
| Condition | Day 1 (ng/mL) | Day 5 (ng/mL) |
| Unstimulated | 399 | 575 |
| Stimulated | 588 | 1091 |
THP-1 human acute monocytic leukemia cells were cultured in RPMI 1640 supplemented with 10% fetal bovine serum and 2 mM L-glutamine. Cells were cultured unstimulated or stimulated with 200 nM PMA until confluent. An aliquot of the cell culture supernate was removed, assayed for human S100A8/S100A9 Heterodimer, and measured 11.6 ng/mL and 38.0 ng/mL, respectively.
Tissue Lysates - Human colon tissue samples were prepared in RIPA buffer with protease
inhibitors. The working buffer was kept on ice and used within 1 day. Aliquots of the tissue
lysates were removed and assayed for human S100A8/S100A9 Heterodimer.
| Sample Type | (ng/mL) |
| Human colon, normal | 14,700 |
| Human colon, cancer | 11,215 |
Fecal Extracts - Three fecal samples were extracted by adding 5 mL extraction buffer (0.1 M Tris, 0.015 M NaCl, 1.0 M Urea, 1.0 mM CaCl2, 0.1 M Citric Acid Monohydrate, 5 mg/mL BSA, and 0.25% Gentamycin Sulfate at pH 8.0) into 100 mg sample. Samples were vortexed for 10 minutes and filtered through a 0.2 μm filter. Samples were assayed for human S100A8/S100A9 Heterodimer and measured 81.2 ng/mL, 8.71 ng/mL, and 1175 ng/mL.
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components.
Cell Culture Supernates, EDTA Plasma, Heparin Plasma, Human Milk, Saliva, Serum, Tissue Lysates, Urine
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (ng/mL) | 5.64 | 12.1 | 25.3 | 6.09 | 12.6 | 24.6 |
| Standard Deviation | 0.155 | 0.370 | 1.14 | 0.352 | 0.606 | 0.792 |
| CV% | 2.7 | 3.1 | 4.5 | 5.8 | 4.8 | 3.2 |
Recovery for Human S100A8/S100A9 Heterodimer Quantikine ELISA Kit
The recovery of human S100A8/S100A9 Heterodimer spiked to levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=4) | 85 | 79-96 |
| Cell Lysis Buffer (n=3) | 94 | 80-104 |
| Human Milk (n=4) | 96 | 83-103 |
| Urine (n=4) | 100 | 91-113 |
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human S100A8/S100A9 Heterodimer were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Scientific Data Images for Human S100A8/S100A9 Heterodimer Quantikine ELISA Kit
Human S100A8/S100A9 Heterodimer ELISA Standard Curve
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: S100A8/S100A9 Heterodimer
S100A8 (also known as MRP8, Calgranulin A, and CP-10) and S100A9 (also known as MRP14 and Calgranulin B) are secreted pro-inflammatory proteins that are upregulated in neutrophils and monocytes at sites of inflammation (e.g. psoriasis, rheumatoid arthritis, cardiac ischemia) and are present at elevated concentrations in rheumatoid arthritis synovial fluid. In the presence of calcium or zinc, S100A8 and S100A9 associate into noncovalent homodimers and heterodimers with each other. The heterodimer additionally binds and sequesters manganese, thereby restricting the growth of Mn-dependent bacteria. The S100A8/A9 heterodimer exhibits functions beyond those performed by the individual proteins. These include binding to fatty acids such as arachidonic acid and promoting astrocyte proliferation. S100A8, S100A9, and the heterodimer each promote neutrophil infiltration into sites of inflammation and inflammatory cytokine production by monocytes.
Long Name
S100 Calcium Binding Protein A8/S100 Calcium Binding Protein A9
Alternate Names
Calprotectin
Entrez Gene IDs
6279 (Human)
Gene Symbol
S100A8
Additional S100A8/S100A9 Heterodimer Products
Product Documents for Human S100A8/S100A9 Heterodimer Quantikine ELISA Kit
Product Specific Notices for Human S100A8/S100A9 Heterodimer Quantikine ELISA Kit
For research use only
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⚠ 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.