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Human MICB DuoSet ELISA

Catalog #: DY1599
19 Citations 1 Review Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
DY1599
Ancillary Products Available
DY008B: DuoSet ELISA Ancillary Reagent Kit 2
DY999B: Substrate Reagent Pack
DY999: Substrate Reagent Pack
Human MICB ELISA Standard Curve
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Product Details
Procedure
Citations (19)
FAQs
Supplemental Products
Reviews (1)
Summary Product Features Kit Content Other Reagents Required Data Examples Product Datasheets Preparation and Storage Background

Human MICB DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
156.0 - 10,000 pg/mL
Sufficient Materials
For fifteen 96-well plates*
Specificity
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 MICB. 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

N/A Human MICB ELISA Standard Curve View Larger

Human MICB ELISA Standard Curve

Product Datasheets

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Product Datasheet
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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.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: MICB

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 B
Entrez Gene IDs:
4277 (Human)
Alternate Names:
MHC class I chain-related protein B; MHC class I mic-B antigen; MHC class I polypeptide-related sequence B; MICB; MIC-B; PERB11.2MHC class I-like molecule PERB11.2-IMX; stress inducible class I homolog

Assay Procedure

GENERAL ELISA PROTOCOL

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 MICB 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.

19 Citations: Showing 1 - 10
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  1. A vaccine targeting resistant tumours by dual T cell plus NK cell attack
    Authors: S Badrinath, MO Dellacheri, A Li, S Zheng, X Zhang, M Sobral, JW Pyrdol, KL Smith, Y Lu, S Haag, H Ijaz, F Connor-Str, T Kaisho, G Dranoff, GC Yuan, DJ Mooney, KW Wucherpfen
    Nature, 2022-05-25;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  2. 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, 2021-07-02;11(1):13754.
    Species: Human
    Sample Types: Cell Lysates
  3. 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, 2019-06-11;5(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. Radiotherapy induces responses of lung cancer to CTLA-4 blockade
    Authors: SC Formenti, NP Rudqvist, E Golden, B Cooper, E Wennerberg, C Lhuillier, C Vanpouille, K Friedman, L Ferrari de, KW Wucherpfen, A Heguy, N Imai, S Gnjatic, RO Emerson, XK Zhou, T Zhang, A Chachoua, S Demaria
    Nat. Med., 2018-11-05;0(0):.
    Species: Human
    Sample Types: Serum
  5. 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, 2017-03-23;0(0):.
    Species: Human
    Sample Types: Serum
  6. Nonblocking Monoclonal Antibody Targeting Soluble MIC Revamps Endogenous Innate and Adaptive Antitumor Responses and Eliminates Primary and Metastatic Tumors.
    Authors: Lu S, Zhang J, Liu D, Li G, Staveley-O'Carroll K, Li Z, Wu J
    Clin Cancer Res, 2015-06-23;21(21):4819-30.
    Species: Mouse
    Sample Types: Serum
  7. 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, 2015-06-12;195(2):736-48.
    Species: Human
    Sample Types: Cell Culture Supernates
  8. 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, 2015-06-09;45(8):2324-34.
    Species: Human
    Sample Types: Plasma
  9. Cytotoxicity and infiltration of human NK cells in in vivo-like tumor spheroids.
    Authors: Giannattasio A, Weil S, Kloess S, Ansari N, Stelzer E, Cerwenka A, Steinle A, Koehl U, Koch J
    BMC Cancer, 2015-05-03;15(0):351.
    Species: Human
    Sample Types: Cell Culture Supernates
  10. 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, 2015-03-16;10(3):e0119961.
    Species: Human
    Sample Types: Peritoneal Fluid
  11. 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, 2013-03-05;21(4):895-903.
    Species: Human
    Sample Types: Serum
  12. 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., 2012-04-19;41(1):83-91.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. Combinatorial Cytotoxicity of Gemcitabine and Cytokine-activated Killer Cells in Hepatocellular Carcinoma via the NKG2D-MICA/B System.
    Authors: Morisaki T, Onishi H, Koya N
    Anticancer Res., 2011-07-01;31(7):2505-10.
    Species: Human
    Sample Types: Cell Culture Supernates
  14. The prognostic significance of soluble NKG2D ligands in B-cell chronic lymphocytic leukemia.
    Authors: Nuckel H, Switala M, Sellmann L, Horn PA, Durig J, Duhrsen U, Kuppers R, Grosse-Wilde H, Rebmann V
    Leukemia, 2010-04-29;24(6):1152-9.
    Species: Human
    Sample Types: Plasma
  15. 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., 2009-11-26;261(2):99-104.
    Species: Human
    Sample Types: Serum
  16. 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., 2009-08-26;106(37):15825-30.
    Species: Human
    Sample Types: Cell Culture Supernates
  17. Obstructing shedding of the immunostimulatory MHC class I chain-related gene B prevents tumor formation.
    Authors: Wu JD, Atteridge CL, Wang X, Seya T, Plymate SR
    Clin. Cancer Res., 2009-01-15;15(2):632-40.
    Species: Mouse
    Sample Types: Cell Lysates
  18. 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, 2009-01-05;113(13):2955-64.
    Species: Human
    Sample Types: Cell Culture Supernates
  19. 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., 2008-06-15;68(12):4746-53.
    Species: Human
    Sample Types: Cell Culture Supernates

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Human MICB DuoSet ELISA
By Anonymous on 06/09/2020
Sample Tested: Serum and Plasma

We used this kit to quantify the MICB in human serum and plasma samples. Works very well and generates a very good standard curve.

Human MICB DuoSet ELISA DY1599