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Human MICA ELISA Standard Curve
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Human MICA DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
96-well strip plate
Sample Volume Required
100 µL
Assay Range
62.5 - 4,000 pg/mL
Sufficient Materials
For fifteen 96-well plates*
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human MICA. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

Normal Goat Serum: (Catalog # DY005)


The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Normal Goat Serum: (Catalog # DY005)


Scientific Data

Human MICA ELISA Standard Curve

Product Datasheets

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Preparation and Storage

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: MICA

MHC class I chain-related genes MICA and MICB are transmembrane glycoproteins that function as ligands for human NKG2D. The two proteins are highly related, sharing 85% amino acid identity, but are also polymorphic. Recognition of MICA or MICB by NKG2D results in the activation of NK cell cytolytic activity and/or cytokine production. MICA/B are minimally expressed on normal cells, but are frequently expressed on or shed from epithelial tumors and can be induced by bacterial and viral infections. MICA and MICB recognition is involved in tumor surveillance, viral infections, and autoimmune diseases.

Long Name:
MHC Class I-related Protein A
Entrez Gene IDs:
100507436 (Human)
Alternate Names:
FLJ60820; MGC111087; MICA; PERB11.1

Assay Procedure


Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent with NGS, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

Citations for Human MICA DuoSet ELISA

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.

30 Citations: Showing 1 - 10
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  1. SARS-CoV-2 escapes direct NK cell killing through Nsp1-mediated downregulation of ligands for NKG2D
    Authors: MJ Lee, MW Leong, A Rustagi, A Beck, L Zeng, S Holmes, LS Qi, CA Blish
    Cell Reports, 2022;41(13):111892.
    Species: Human
    Sample Types: Cell Culture Supernates
  2. Bronchoalveolar lavage fluid reveals factors contributing to the efficacy of PD-1 blockade in lung cancer
    Authors: K Masuhiro, M Tamiya, K Fujimoto, S Koyama, Y Naito, A Osa, T Hirai, H Suzuki, N Okamoto, T Shiroyama, K Nishino, Y Adachi, T Nii, Y Kinugasa-K, A Kajihara, T Morita, S Imoto, S Uematsu, T Irie, D Okuzaki, T Aoshi, Y Takeda, T Kumagai, T Hirashima, A Kumanogoh
    JCI Insight, 2022;7(9):.
    Species: Human
    Sample Types: BALF
  3. MICA*019 Allele and Soluble MICA as Biomarkers for Ankylosing Spondylitis in Taiwanese
    Authors: CM Wang, KP Tan, YJ Jan Wu, JC Lin, JW Zheng, AL Yu, JM Wu, JY Chen
    Journal of personalized medicine, 2021;11(6):.
    Species: Human
    Sample Types: Serum
  4. Cord blood and amniotic membrane extract eye drop preparations display immune-suppressive and regenerative properties
    Authors: D Samarkanov, S Cox, D Hernandez, L Rodriguez, ML Pérez, A Madrigal, A Vilarrodon, S Querol, RP Casaroli-M
    Scientific Reports, 2021;11(1):13754.
    Species: Human
    Sample Types: Cell Lysates
  5. Increase of Antitumoral Effects of Cytokine-Induced Killer Cells by Antibody-Mediated Inhibition of MICA Shedding
    Authors: X Wu, Y Zhang, Y Li, IGH Schmidt-Wo
    Cancers (Basel), 2020;12(7):.
    Species: Human
    Sample Types: Cell Culture Supernates
  6. Targetable mechanisms driving immunoevasion of persistent senescent cells link chemotherapy-resistant cancer to aging
    Authors: DP Muñoz, SM Yannone, A Daemen, Y Sun, F Vakar-Lope, M Kawahara, AM Freund, F Rodier, JD Wu, PY Desprez, DH Raulet, PS Nelson, LJ van 't Vee, J Campisi, JP Coppé
    JCI Insight, 2019;5(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. Race-associated expression of MHC class I polypeptide-related sequence A (MICA) in prostate cancer
    Authors: MJ Sakiyama, I Espinoza, A Reddy, F de Carlo, A Kumar, AS Levenson, S Bae, X Zhou, PP Claudio, J Lewin, V Manucha, CR Pound, S Vijayakuma, CR Gomez
    Exp. Mol. Pathol., 2019;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  8. Persistently elevated soluble MHC class I polypeptide-related sequence A and transforming growth factor-?1 levels are poor prognostic factors in head and neck squamous cell carcinoma after definitive chemoradiotherapy
    Authors: JL Chen, CC Chang, YS Huang, HY Kuo, TY Chen, CW Wang, SH Kuo, YL Lin
    PLoS ONE, 2018;13(8):e0202224.
    Species: Human
    Sample Types: Plasma
  9. MHC class I chain-related protein A and B (MICA and MICB) are predominantly expressed intracellularly in tumour and normal tissue
    Authors: H Ghadially, L Brown, C Lloyd, L Lewis, A Lewis, J Dillon, R Sainson, J Jovanovic, NJ Tigue, D Bannister, L Bamber, V Valge-Arch, RW Wilkinson
    Br. J. Cancer, 2017;0(0):.
    Species: Human
    Sample Types: Serum
  10. All-trans retinoic acid enhances cytotoxicity of CIK cells against human lung adenocarcinoma by upregulating MICA and IL-2 secretion
    Authors: XY Fan, PY Wang, C Zhang, YL Zhang, Y Fu, C Zhang, QX Li, JN Zhou, BE Shan, DW He
    Sci Rep, 2017;7(1):16481.
  11. SEP enhanced the antitumor activity of 5-fluorouracil by up-regulating NKG2D/MICA and reversed immune suppression via inhibiting ROS and caspase-3 in mice
    Authors: M Ke, H Wang, Y Zhou, J Li, Y Liu, M Zhang, J Dou, T Xi, B Shen, C Zhou
    Oncotarget, 2016;7(31):49509-49526.
    Species: Human
    Sample Types: Cell Culture Supernates
  12. Genetics Variants and Serum Levels of MHC Class I Chain-related A in Predicting Hepatocellular Carcinoma Development in Chronic Hepatitis C Patients Post Antiviral Treatment
    EBioMedicine, 2016;0(0):.
    Species: Human
    Sample Types: Serum
  13. Genotoxic Stress Induces Senescence-Associated ADAM10-Dependent Release of NKG2D MIC Ligands in Multiple Myeloma Cells.
    Authors: Zingoni A, Cecere F, Vulpis E, Fionda C, Molfetta R, Soriani A, Petrucci M, Ricciardi M, Fuerst D, Amendola M, Mytilineos J, Cerboni C, Paolini R, Cippitelli M, Santoni A
    J Immunol, 2015;195(2):736-48.
  14. CEACAM1-3S Drives Melanoma Cells into NK Cell-Mediated Cytolysis and Enhances Patient Survival.
    Authors: Ullrich N, Heinemann A, Nilewski E, Scheffrahn I, Klode J, Scherag A, Schadendorf D, Singer B, Helfrich I
    Cancer Res, 2015;75(9):1897-907.
    Species: Human
    Sample Types: Cell Culture Supernates
  15. Umbilical cord blood plasma contains soluble NKG2D ligands that mediate loss of natural killer cell function and cytotoxicity.
    Authors: Cox S, Laza-Briviesca R, Pearson H, Soria B, Gibson D, Gomez S, Madrigal J, Saudemont A
    Eur J Immunol, 2015;45(8):2324-34.
    Species: Human
    Sample Types: Plasma
  16. Soluble ligands for the NKG2D receptor are released during endometriosis and correlate with disease severity.
    Authors: Gonzalez-Foruria I, Santulli P, Chouzenoux S, Carmona F, Batteux F, Chapron C
    PLoS ONE, 2015;10(3):e0119961.
    Species: Human
    Sample Types: Peritoneal Fluid
  17. The MICA-129 dimorphism affects NKG2D signaling and outcome of hematopoietic stem cell transplantation
    EMBO Mol Med, 2015;7(11):1480-502.
    Species: Human
    Sample Types: Cell Culture Supernates
  18. TLR ligands stimulation protects MSC from NK killing.
    Authors: Giuliani M, Bennaceur-Griscelli A, Nanbakhsh A, Oudrhiri N, Chouaib S, Azzarone B, Durrbach A, Lataillade J
    Stem Cells, 2014;32(1):290-300.
    Species: Human
    Sample Types: Cell Culture Supernates
  19. Expression of major histocompatibility complex class I-related chain A/B (MICA/B) in pancreatic carcinoma.
    Authors: Dambrauskas Z, Svensson H, Joshi M, Hyltander A, Naredi P, Iresjo B
    Int J Oncol, 2014;44(1):99-104.
    Species: Human
    Sample Types: Serum
  20. Rescue of impaired NK cell activity in hodgkin lymphoma with bispecific antibodies in vitro and in patients.
    Authors: Reiners K, Kessler J, Sauer M, Rothe A, Hansen H, Reusch U, Hucke C, Kohl U, Durkop H, Engert A, von Strandmann E
    Mol Ther, 2013;21(4):895-903.
    Species: Human
    Sample Types: Serum
  21. Soluble MICA and a MICA variation as possible prognostic biomarkers for HBV-induced hepatocellular carcinoma.
    Authors: Kumar V, Yi Lo P, Sawai H, Kato N, Takahashi A, Deng Z, Urabe Y, Mbarek H, Tokunaga K, Tanaka Y, Sugiyama M, Mizokami M, Muroyama R, Tateishi R, Omata M, Koike K, Tanikawa C, Kamatani N, Kubo M, Nakamura Y, Matsuda K
    PLoS ONE, 2012;7(9):e44743.
    Species: Human
    Sample Types: Serum
  22. Valproic acid cooperates with hydralazine to augment the susceptibility of human osteosarcoma cells to Fas- and NK cell-mediated cell death.
    Authors: Yamanegi K, Yamane J, Kobayashi K
    Int. J. Oncol., 2012;41(1):83-91.
    Species: Human
    Sample Types: Cell Culture Supernates
  23. Genome-wide association study identifies a susceptibility locus for HCV-induced hepatocellular carcinoma.
    Authors: Kumar V, Kato N, Urabe Y, Takahashi A, Muroyama R, Hosono N, Otsuka M, Tateishi R, Omata M, Nakagawa H, Koike K, Kamatani N, Kubo M, Nakamura Y, Matsuda K
    Nat. Genet., 2011;43(5):455-8.
    Species: Human
    Sample Types: Serum
  24. Human breast cancer cells enhance self tolerance by promoting evasion from NK cell antitumor immunity.
    Authors: Mamessier E, Sylvain A, Thibult ML, Houvenaeghel G, Jacquemier J, Castellano R, Goncalves A, Andre P, Romagne F, Thibault G, Viens P, Birnbaum D, Bertucci F, Moretta A, Olive D
    J. Clin. Invest., 2011;121(9):3609-22.
    Species: Human
    Sample Types: Cell Culture Supernates
  25. The role of activation-induced cell death in the higher onset of spontaneous apoptosis of NK cell subsets in patients with metastatic epithelial cancer.
    Authors: Stanzer S, Janesch B, Resel M, Augustin T, Samonigg H, Bauernhofer T
    Cell. Immunol., 2010;261(2):99-104.
    Species: Human
    Sample Types: Serum
  26. Inhibition of NKG2D expression in NK cells by cytokines secreted in response to human cytomegalovirus infection.
    Authors: Muntasell A, Magri G, Pende D, Angulo A, Lopez-Botet M
    Blood, 2010;115(25):5170-9.
    Species: Human
    Sample Types: Cell Culture Supernates
  27. Detuning CD8+ T lymphocytes by down-regulation of the activating receptor NKG2D: role of NKG2D ligands released by activated T cells.
    Authors: Cerboni C, Ardolino M, Santoni A, Zingoni A
    Blood, 2009;113(13):2955-64.
    Species: Human
    Sample Types: Cell Culture Supernates
  28. Biologic activity of irradiated, autologous, GM-CSF-secreting leukemia cell vaccines early after allogeneic stem cell transplantation.
    Authors: Ho VT, Vanneman M, Kim H, Sasada T, Kang YJ, Pasek M, Cutler C, Koreth J, Alyea E, Sarantopoulos S, Antin JH, Ritz J, Canning C, Kutok J, Mihm MC, Dranoff G, Soiffer R
    Proc. Natl. Acad. Sci. U.S.A., 2009;106(37):15825-30.
    Species: Human
    Sample Types: Cell Culture Supernates
  29. Clinical and immunological evaluation of zoledronate-activated Vgamma9gammadelta T-cell-based immunotherapy for patients with multiple myeloma.
    Authors: Abe Y, Muto M, Nieda M, Nakagawa Y, Nicol A, Kaneko T, Goto S, Yokokawa K, Suzuki K
    Exp. Hematol., 2009;37(8):956-68.
    Species: Human
    Sample Types: Serum
  30. Hypoxia increases tumor cell shedding of MHC class I chain-related molecule: role of nitric oxide.
    Authors: Siemens DR, Hu N, Sheikhi AK, Chung E, Frederiksen LJ, Pross H, Graham CH
    Cancer Res., 2008;68(12):4746-53.
    Species: Human
    Sample Types: Cell Culture Supernates


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By Anonymous on 03/27/2018
Sample Tested: Serum and Plasma

Highly recommend using this DuoSet along the ancillary kit 2, otherwise may encounter some issues regarding BSA quality, personally used the ancillary kit then all issues were resolved