Macrophage inflammatory protein-1 alpha(MIP-1 alpha), also called CCL3, LD78. The cDNA for MIP-1 alpha predicts a mature peptide of 69aa with a molecular mass of 7,889 daltons. LD78 is a member of a newly identified superfamily of small inducible proteins involved in inflammatory responses, wound healing and tumorigenesis. MIP-1 alpha is a chemokine that has pro-inflammatory and stem cell inhibitory activities in vitro. It constitutes an important second signal for mast cell degranulation in the conjunctiva in vivo and consequently for acute-phase disease.
Sample Type
Cell culture supernates, serum and plasma (heparin, EDTA, citrate).
Intended Use
For quantitative detection of rat MIP-1 alpha in cell culture supernates, serum and plasma (heparin, EDTA, citrate).
Specificity
Natural and recombinant rat MIP-1 alpha
Test Principle
This rat MIP-1 alpha BioAssay™ ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for MIP-1 alpha has been precoated onto 96-well plates. Standards(E.coli) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for MIP-1 alpha is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was 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 rat MIP-1 alpha amount of sample captured in plate.
Kit Components
96-well plate precoated with rat MIP-1 alpha antibody, 1 Lyophilized recombinant rat MIP-1 alpha standard, 10ng/tube×2 Biotinylated rat MIP-1 alpha antibody, 130ul (dilution 1:100) Avidin-Biotin-Peroxidase Complex (ABC), 130ul (dilution 1:100) Sample diluent buffer, 30ml Antibody diluent buffer, 12ml ABC diluent buffer, 12ml TMB color developing agent, 10ml TMB stop solution, 10ml
Storage and Stability
Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Important Note
This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.