Human AMICA / JAML ELISA Kit

Catalog No.: BEK1291

Size: 96T

Range: 62.5 pg/ml - 4000 pg/ml

Sensitivity < 10 pg/ml

Storage and Expiration: Store at 2-8℃ for 6 months, or at -20℃ for 12 months. 

Application: For quantitative detection of AMICA / JAML in human serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Introduction

JAML or Junctional Adhesion Molecule-Like, or AMICA1 is a JAM transmembrane protein family member. It is composed of two extracellular immunoglobulin-like domains, a membrane-spanning region, and a cytoplasmic tail involved in activation signaling. A known ligand of JAML is Coxsackie virus and Adenovirus Receptor (CXADR in humans and CAR in mice) which has been shown to localize to the tight junctions of epithelial cells. JAML-mediated activation of CAR is required for neutrophil extravasation in addition to other leukocyte/epithelial cell interaction models. Other members of the JAM family of transmembrane proteins include JAM1, JAM2 and JAM3.

Principle of the Assay  

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

Kit components

1. One 96-well plate pre-coated with anti-human AMICA / JAML antibody
2. Lyophilized human AMICA / JAML standards: 2 tubes (10ng / tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-human AMICA / JAML antibody (Concentrated): 130μl. Dilution: 1:100
5. Antibody diluent buffer: 12ml
6. Avidin-Biotin-Peroxidase Complex (ABC) (Concentrated): 130μl. Dilution: 1:100
7. ABC diluent buffer: 12ml
8. TMB substrate: 10ml
9. Stop solution: 10ml
10. Wash buffer (25X): 30ml

Note: Reconstitute standards and test samples with Kit Component 3.

Material Required But Not Provided

1. 37℃ 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