Total SARS-COV-2 IgG

Human MICA ELISA Assay

$635.00

The Human MICA ELISA Assay is for the in-vitro quantitative determination of MHC class I chain-related gene A glycoprotein, in cell culture supernatants and buffered solutions.

NOTE Serum and Plasma quantification: high detection levels are found in human serum and plasma samples. The appearance of a high signal can be the result of the matrix and/or interaction with other molecules. Consequently we do not recommend the use of serum and plasma and the use of such samples with this kit is solely at the discretion of the user.

The Eagle Biosciences Human MICA ELISA Assay kit is for Research Use Only.

SKU: MIC31-K01 Categories: , ,

Human MICA ELISA Assay

The Human MICA ELISA Assay is For Research Use Only

Size: 1×96 wells
Sensitivity: 123 pg/ml
Dynamic Range: 156 – 5000 pg/ml
Incubation Time: 3h 50 min
Sample Type: Supernatant, Buffered Solutions
Sample Size: 100 µl

Assay Background

MICA is a transmembrane glycoprotein that functions as a ligand for human NKG2D, an activating receptor expressed on NK Cells, NKT Cells, dg T-cells and CD8+ba T-cells. Recognition of MICA by NKG2D results in the activation of cytolytic activity and/or cytokine production by these effectors cells. MICA recognition is involved in tumour surveillance, viral infections, and autoimmune diseases.

Major histocompatibility complex (MHC) class I chain-related gene A and B (MICA and MICB) are transmembrane glycoproteins that function as ligand for NKG2D. These two proteins possess three extracellular immunoglobulin-like domains, but have no capacity to bind peptide or interact with b2-microglobulin. The genes encoding MICA/B are found within the MHC on human chromosome 6.

MICA and MICB have no role in antigen presentation but function as signal of cellular distress and interact with NKG2D-DAP10, the activating receptor. They are frequently expressed in epithelial tumour and may promote anti tumour NK and T-cell response.

Intestinal cells express MICA/MICB which are up-regulated under stress and in many gastrointestinal tumours. Release of MIC molecules from the cell surface is thought to constitute in immune escape mechanism of tumour cells.

MICA/MICB expression is elevated in the sera of patients with colorectal carcinoma and widely expressed in prostate carcinoma : MICA/MICB may be a novel biomarker for prostate cancer and its expression is also used as a monitor in Crohn’s disease.

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Additional Information

Assay Principle

The Eagle Biosciences MICA Kit is a solid phase sandwich Enzyme Linked-Immuno-Sorbent Assay (ELISA). A monoclonal antibody specific for MICA has been coated onto the wells of the microtiter strips provided. Samples, including standards of known MICA concentrations and unknown are pipetted into these wells.
During the first incubation, the MICA antigen is added to wells. After washing, a biotinylated monoclonal antibody specific for MICA is incubated. Then the enzyme (streptavidin-horse radish peroxidase) is added. After incubation and washing to remove all unbound enzyme, a substrate solution of the bound enzyme is added to induce a coloured reaction product. The intensity of this coloured product is directly proportional to the concentration of MICA present in the samples.

  1. Add 100µl of each, Sample and zero (appropriate standard diluent) in duplicate to appropriate number of wells
  2. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) for 2 hour(s)
  3. Remove the cover and wash the plate
  4. Add 50µl of diluted biotinylated anti-MICA to all wells
  5. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) for 1 hour(s)
  6. Repeat wash step 3.
  7. Add 100μl of Streptavidin-HRP solution into all wells
  8. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) for 30 min
  9. Repeat wash step 3.
  10. Add 100μl of ready-to-use TMB Substrate Solution into all wells
  11. Incubate in the dark for 5-15 minutes* at room temperature. Avoid direct exposure to light by wrapping the plate in aluminium foil.
  12. Add 100μl of H2SO4:Stop Reagent into all wells

Typical Standard Curve

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