MICA ELISA Kit

Catalog No. ABIN1112748

Product Details MICA ELISA Kit

Target: MICA (MHC Class I Polypeptide-Related Sequence A (MICA))

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 62.5-4000 pg/mL

Minimum Detection Limit: 62.5 pg/mL

Application: ELISA

Analytical Method: Quantitative

Sensitivity: < 10 pg/mL

Components: 1. One 96-well plate pre-coated with anti-Human MICA antibody 2. Lyophilized Human MICA standards: 2 tubes (10ng / tube) 3. Sample / Standard diluent buffer: 30ml 4. Biotin conjugated anti-Human MICA antibody (Concentrated): 130 µl.

Material not included: 1. 37 °C incubator 2. Microplate reader (wavelength: 450nm) 3. Precise pipette and disposable pipette tips 4. Automated plate washer 5. ELISA shaker 6. 1.5ml of Eppendorf tubes 7. Plate cover 8. Absorbent filter papers 9. Plastic or glass container with volume of above 1L

Application Details

Comment:

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-MICA polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-MICA polyclonal antibody was used as detection antibodies. The standards test samples and biotin conjugated detection antibody were added - the wells subsequently and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were used - visualize HRP enzymatic reaction. TMB was catalyzed by HRP - produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional - the MICA amount of sample captured in plate. Read the O.D. absorbance at 450 nm in a microplate reader and then the concentration of MICA can be calculated.

Plate: Pre-coated

Reagent Preparation:

1. Before the experiment, centrifuge each kit component for several minutes to bring down all reagents to the bottom of tubes. 2. It is recommend to measure each standard and sample in duplicate. 3. Do NOT let the plate completely dry at any time! Since the dry condition can inactivate the biological material on the plate. 4. Do not reuse pipette tips and tubes to avoid cross contamination. 5. Do not use the expired components and the components from different batches. 6. To avoid the marginal effect of plate incubation for temperature differences (the marginal wells always get stronger reaction), it is recommend to equilibrate the ABC working solution and TMB substrate for at least 30 min at room temperature (37°C ) before adding to wells.The TMB substrate (Kit Component 8) is colorless and transparent before use, if not, please contact us for replacement.

Sample Preparation:

Preparation of sample and reagents 1. Sample Isolate the test samples soon after collecting, then, analyze immediately (within 2 hours). Or aliquot and store at -20 °C for long term. Avoid multiple freeze-thaw cycles.
Tissue lysates and cell culture supernatants: Centrifuge to remove precipitate, analyze immediately or aliquot and store at -20 °C .
Serum: Coagulate the serum at room temperature (about 4 hours). Centrifuge at approximately 1000 × g for 15 min. Analyze the serum immediately or aliquot and store at -20 °C .
Plasma: Collect plasma with heparin or EDTA as the anticoagulant. Centrifuge for 15 min at 1000 x g within 30 min of collection. For eliminating the platelet effect, suggesting that further centrifugation for 10 min at 2-8°C at 10000 x g. Analyze immediately or aliquot and store frozen at -20 °C. Citrate can not be used as anticoagulant here. Note: 1. Coagulate blood samples completely, then, centrifuge, and avoid hemolysis and particle. 2. NaN3 can not be used as test sample preservative, since it is the inhibitor for HRP.

Restrictions: For Research Use only

Handling

Preservative: Sodium azide, Thimerosal (Merthiolate)

Target Details for MICA

Target: MICA (MHC Class I Polypeptide-Related Sequence A (MICA))

Alternative Name: MICA (MICA Products)

Synonyms: MIC-A ELISA Kit, PERB11.1 ELISA Kit, MHC class I polypeptide-related sequence A ELISA Kit, MICA ELISA Kit

Background: MHC class I polypeptide-related sequence A(MICA) is a protein that in humans is encoded by the MICA gene. The MICA gene is located near HLA-B on chromosome 6. Nalabolu et al. (1996) showed that the MICA and MICB (602436) genes occur in a 200-kb region spanning the TNFA (191160) and TNFB (153440) cluster at chromosome 6p21.3. The protein product is expressed on the cell surface, although unlike canonical class I molecules does not seem to associate with beta-2-microglobulin. It is thought that MICA functions as a stress-induced antigen that is broadly recognized by NK cells, NKT cells, and most of the subtypes of T cells. MICA is the ligand for NK cell activating receptor NKG2D.

Pathways: Activation of Innate immune Response, Transition Metal Ion Homeostasis