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Human S100A8/S100A9 Heterodimer Quantikine ELISA Kit

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Human S100A8/S100A9 Heterodimer ELISA Standard Curve
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Summary Sample Values Precision Recovery Linearity Data Examples Product Datasheets Preparation and Storage Background Related Research Areas

Human S100A8/S100A9 Heterodimer Quantikine ELISA Kit Summary

Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Tissue Lysates (50 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL), Saliva (10 uL), Urine (50 uL), Human Milk (10 uL), Fecal Extract (50 uL)
Sensitivity
0.215 ng/mL
Assay Range
0.6 - 40 ng/mL (Cell Culture Supernates, Tissue Lysates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine, Human Milk)
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.

Sample TypeMean (ng/mL)Range (ng/mL)Standard Deviation (ng/mL)
Serum (n=72)2015481-65401219
EDTA plasma (n=40)473127-1395295
Heparin plasma (n=40)830298-1640314
Saliva (n=10)72712137-18,9605336
Urine (n=10)1020.858-35999.7
Human milk (n=10)130834.0-57201876

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.

ConditionDay 1 (ng/mL)Day 5 (ng/mL)
Unstimulated399575
Stimulated5881091

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, normal14,700
Human colon, cancer11,215

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.



Product Summary

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.

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, Tissue Lysates, 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 (ng/mL) 5.64 12.1 25.3 6.09 12.6 24.6
Standard Deviation 0.155 0.37 1.14 0.352 0.606 0.792
CV% 2.7 3.1 4.5 5.8 4.8 3.2

Recovery

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.
Human S100A8/S100A9 Heterodimer ELISA Linearity

Scientific Data

N/A Human S100A8/S100A9 Heterodimer ELISA Standard Curve View Larger

Human S100A8/S100A9 Heterodimer ELISA Standard Curve

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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: S100A8/S100A9 Heterodimer

S100A8 (also MRP8 and calgranulin A) is a 10 kDa member of the S100 family, EF-hand superfamily of Ca-binding proteins. It is produced by neutrophils and monocytes, and forms Ca2+-dependent heterodimer/heterotetramer complexes (termed calprotectin) with S100A9. It functions both intracellularly and extracellularly, where it binds to RAGE and CD36. Human S100A8 is 93 amino acids (aa) in length. It contains two EF-hand motifs (aa 12 - 47 and 46 - 81) and one high-affinity Ca2+-binding site (aa 59 - 70). There may be one splice form that shows a 15 aa substitution for the C-terminal 14 amino acids. Although mouse S100A8 is cleaved by MMP-2 after Asn21, it is unclear if human S100A8 is susceptible. Full-length human S100A8 is 57% and 74% aa identical to mouse and canine S100A8, respectively.

Long Name:
S100 Calcium Binding Protein A8/S100 Calcium Binding Protein A9
Entrez Gene IDs:
6279 (Human)
Alternate Names:
60B8AG; CAGA; calgranulin A; Calprotectin L1L subunit; Calprotectin; CFAG; CGLA; CP-10; Cystic fibrosis antigen; L1AG; Leukocyte L1 complex light chain; MA387; MIF; Migration inhibitory factor-related protein 8; MRP8; NIF; p8; protein S100-A8; S100-A8; S100A8/S100A9 Heterodimer; Urinary stone protein band A
&#9888; 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 S100A8/S100A9 Heterodimer Quantikine ELISA Kit

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

15 Citations: Showing 1 - 10
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  1. Performance of 11 Host Biomarkers Alone or in Combination in the Diagnosis of Late-Onset Sepsis in Hospitalized Neonates: The Prospective EMERAUDE Study
    Authors: Pons, S;Trouillet-Assant, S;Subtil, F;Abbas-Chorfa, F;Cornaton, E;Berthiot, A;Galletti, S;Plat, A;Rapin, S;Trapes, L;Generenaz, L;Brengel-Pesce, K;Callies, A;Plaisant, F;Claris, O;Portefaix, A;Flamant, C;Butin, M;
    Biomedicines
    Species: Human
    Sample Types: Serum
  2. S100A8-mediated metabolic adaptation controls HIV-1 persistence in macrophages in vivo
    Authors: F Real, A Zhu, B Huang, A Belmellat, A Sennepin, T Vogl, C Ransy, M Revol, R Arrigucci, A Lombès, J Roth, ML Gennaro, F Bouillaud, S Cristofari, M Bomsel
    Nature Communications, 2022-10-11;13(1):5956.
    Species: Human
    Sample Types: Tissue Homogenates
  3. ESR1 mutant breast cancers show elevated basal cytokeratins and immune activation
    Authors: Z Li, O McGinn, Y Wu, A Bahreini, NM Priedigkei, K Ding, S Onkar, C Lampenfeld, CA Sartorius, L Miller, M Rosenzweig, O Cohen, N Wagle, JK Richer, WJ Muller, L Buluwela, S Ali, TC Bruno, DAA Vignali, Y Fang, L Zhu, GC Tseng, J Gertz, JM Atkinson, AV Lee, S Oesterreic
    Nature Communications, 2022-04-19;13(1):2011.
    Species: Human
    Sample Types: Plasma
  4. Core genes involved in the regulation of acute lung injury and their association with COVID-19 and tumor progression: A bioinformatics and experimental study
    Authors: AV Sen'kova, IA Savin, EV Brenner, MA Zenkova, AV Markov
    PLoS ONE, 2021-11-22;16(11):e0260450.
    Species: Human
    Sample Types: Plasma
  5. An Intervention With Michigan-Grown Wheat in Healthy Adult Humans to Determine Effect on Gut Microbiota: Protocol for a Crossover Trial
    Authors: GA Kinney, EN Haddad, LS Garrow, PKW Ng, SS Comstock
    JMIR research protocols, 2021-10-06;10(10):e29046.
    Species: Human
    Sample Types: Stool
  6. Differential Regulation of Circulating Soluble Receptor for Advanced Glycation End Products (sRAGEs) and Its Ligands S100A8/A9 Four Weeks Post an Exercise Intervention in a Cohort of Young Army Recruits
    Authors: IA Drosatos, JN Tsoporis, S Izhar, S Gupta, G Tsirebolos, E Sakadakis, AS Triantafyl, A Rigopoulos, D Rigopoulos, LS Rallidis, I Rizos, TG Parker
    Biomolecules, 2021-09-13;11(9):.
    Species: Human
    Sample Types: Plasma
  7. Biomarkers of systemic inflammation, soluble IL-2Ralpha and the multiple sclerosis-associated IL2RA SNP rs2104286 in healthy subjects and multiple sclerosis patients
    Authors: S Buhelt, HB Søndergaar, MR Mahler, S Cobanovic, L Börnsen, C Ammitzbøll, AB Oturai, F Sellebjerg
    Multiple sclerosis and related disorders, 2021-07-07;54(0):103140.
    Species: Human
    Sample Types: Serum
  8. Circulating levels of tight junction proteins in multiple sclerosis: Association with inflammation and disease activity before and after disease modifying therapy
    Authors: A Olsson, S Gustavsen, AR Langkilde, TH Hansen, F Sellebjerg, H Bach Sønde, AB Oturai
    Multiple sclerosis and related disorders, 2021-07-03;54(0):103136.
    Species: Human
    Sample Types: Serum
  9. Biomarkers of inflammation and epithelial barrier function in multiple sclerosis
    Authors: A Olsson, S Gustavsen, IC Hasselbalc, AR Langkilde, F Sellebjerg, AB Oturai, HB Søndergaar
    Mult Scler Relat Disord, 2020-09-19;46(0):102520.
    Species: Human
    Sample Types: Serum
  10. Toll-like receptor involvement in adolescent scoliotic facet joint degeneration
    Authors: DG Bisson, K Sheng, S Kocabas, E Krock, A Teles, N Saran, JA Ouellet, L Haglund
    J. Cell. Mol. Med., 2020-08-27;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  11. Normative values for circulating intestinal fatty acid binding protein and calprotectin across gestational ages
    Authors: DR Shores, J Fundora, M Go, F Shakeel, S Brooks, SM Alaish, J Yang, CP Sodhi, DJ Hackam, A Everett
    BMC Pediatr, 2020-05-26;20(1):250.
    Species: Human
    Sample Types: Serum
  12. Body Mass Index and Calprotectin Blood Level Correlation in Healthy Children: An Individual Patient Data Meta-Analysis
    Authors: A Grand, E Rochette, F Dutheil, D Gozal, V Calcaterra, R Berni Cana, N Cobanoglu, JPM Derikx, G Terrin, B Pereira, E Merlin
    J Clin Med, 2020-03-20;9(3):.
    Species: Human
    Sample Types: Plasma
  13. Activation of a TLR9 mediated innate immune response in preeclampsia
    Authors: RD Williamson, FP McCarthy, LC Kenny, CM McCarthy
    Sci Rep, 2019-04-11;9(1):5920.
    Species: Human
    Sample Types: Plasma
  14. The rationale and design of the personal diet study, a randomized clinical trial evaluating a personalized approach to weight loss in individuals with pre-diabetes and early-stage type 2 diabetes
    Authors: CJ Popp, DE St-Jules, L Hu, L Ganguzza, P Illiano, M Curran, H Li, A Schoenthal, M Bergman, AM Schmidt, E Segal, A Godneva, MA Sevick
    Contemp Clin Trials, 2019-03-04;0(0):.
    Species: Human
    Sample Types:
  15. Dietary zinc alters the microbiota and decreases resistance to Clostridium difficile infection
    Nat Med, 2016-09-26;0(0):.
    Species: Human
    Sample Types: Serum

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Reviews for Human S100A8/S100A9 Heterodimer Quantikine ELISA Kit

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Human S100A8/S100A9 Heterodimer Quantikine ELISA Kit
By Mike Ehrhardt on 11/24/2020
Sample Tested: EDTA Plasma

This ELISA performed as expected with low % CV's

Human S100A8/S100A9 Heterodimer Quantikine ELISA Kit DS8900
Human S100A8/S100A9 Heterodimer Quantikine ELISA Kit
By Anonymous on 09/07/2017
Sample Tested: Human cells