Macrophage Inflammatory Proteins (MIP) belong to the family of chemotactic cytokines known as chemokines. In humans, there are two major forms, MIP-1alpha and MIP-1beta that are now officially named CCL3 and CCL4 respectively. Both are major factors produced by macrophages after they are stimulated with bacterial endotoxins. They activate human granulocytes (neutrophils, eosinophils and basophils) which can lead to acute neutrophilic inflammation. They also induce the synthesis and release of other pro-inflammatory cytokines such as interleukin 1 (IL-1), IL-6 and TNF-alpha from fibroblasts and macrophages. The genes for CCL3 and CCL4 are both located on human chromosome 17.
Macrophage Inflammatory Protein-1 alpha (MIP-1a) Human Recombinant produced in E. coli is a single, non-glycosylated, polypeptide chain containing 70 amino acids and having a molecular mass of 7820 Dalton. The MIP-1a is purified by proprietary chromatographic techniques. The protein was lyophilized from 0.55 mg/ml solution containing no additives. Purity is greater than 98.0% as determined by RP-HPLC and SDS-PAGE. The biological activity is calculated by the ability of chemo-attraction of Human monocytes using 1-10 ng/ml. The sequence of the first five N-terminal amino acids was determined and was found to be, Ala-Ser-Leu-Ala-Ala.
Protein quantitation was carried out by two independent methods:
1. UV spectroscopy at 280 nm using the absorbency value of 0.9 as the extinction coefficient for a 0.1% (1mg/ml) solution. This value is calculated by the PC GENE computer analysis program of protein sequences (IntelliGenetics).
2. Analysis by RP-HPLC, using a calibrated solution of MIP-1a as a Reference Standard.
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