Certificate of Analysis and Data Sheet

Source:
E.Coli

Catalog No.
CYT-310

Background:

MIA acts as a potent tumor cell growth inhibitor for malignant melanoma cells and some other neuroectodermal tumors, including gliomas, in an autocrine fashion. In a study of human melanoma cell lines with different metastatic capacity MIA mRNA expression appeared to be inversely correlated with pigmentation. MIA has been shown to represent a very sensitive and specific serum marker for systemic malignant melanoma that might be useful for staging of primary melanomas, detection of progression from localized to metastatic disease during follow-up, and monitoring therapy of advanced melanomas.

Description :

Recombinant Human MIA produced in E.Coli is a single, non-glycosylated, polypeptide chain consisting of 108 amino having a total molecular mass of 12237 Dalton. The Melanoma Inhibitory protein (MIA) was identified as an inhibitor of in vitro growth of malignant melanoma cells. The protein contains a SH3 domain. Human Melanoma Inhibitory protein is purified by proprietary chromatographic techniques.

Physical Appearance:

Sterile Filtered White lyophilized (freeze-dried) powder.

Formulation:

Recombinant MIA was lyophilized from a concentrated (1mg/ml) solution containing 20mM Potassium-phosphate pH=7 and 150mM potassium chloride.

Solubility:

It is recommended to reconstitute the lyophilized Human MIA in sterile 18MO-cm H2O not less than 100ug/ml, which can then be further diluted to other aqueous solutions.

Stability:

Lyophilized Recombinant MIA although stable at room temperature for 3 weeks, should be stored desiccated below -18 C. Upon reconstitution Recombinant MIA should be stored at 4 C between 2-7 days and for future use below -18 C. For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA).
Please avoid freeze-thaw cycles.

Purity:

Greater than 95.0% as determined by:
(a) Analysis by RP-HPLC.
(b) Anion-exchange FPLC.
(c) Analysis by reducing and non-reducing SDS-PAGE Silver Stained gel.

Amino acid sequence:

Agrees with the sequence of native human MIA with an addition N-terminal Methionine residue. MGPMPKLADRKLCADQECSSHPISMAVALQDYMAPDCRFLTIHRGQVVYVFSLKGRGRFLWGGSVQGDYYGDLAARLGYFPSSIVREDQTLKVDVKTDKWDFYCQ

Dimers and aggregates:

Less than 1% as determined by silver-stained SDS-PAGE gel analysis.

Protein content:

Protein quantitation was carried out by two independent methods:
1. UV spectroscopy at 280 nm using the absorbption coefficient of 19300 M^-1cm^-1 .
2. Analysis by RP-HPLC, using a calibrated solution of MIA as a Reference Standard.

Endotoxin:

Less than 0.1 ng/ug (IEU/ug) of Human MIA.

Usage:

This material is offered by ProSpec-TechnoGene for research, laboratory or further evaluation purposes.

Gene:

Name:MIA

Protein synonyms/aliases:

Melanoma derived growth regulatory protein precursor (Melanomainhibitory activity).

Protein Family:

Belongs to the MIA/OTOR family.

Protein Domains:

Domains:
•IPR011511   Variant SH3
•IPR001452   Src homology-3

Protein links:

Precursor- Protein structure and amino acid sequence:

Latest Publications:

1. Solution structure and dynamics of melanoma inhibitory activity protein.
J Biomol NMR 2002 Mar;22(3):211-23
2. Structure of melanoma inhibitory activity protein, a member of a recently identified family of secreted proteins.
Proc Natl Acad Sci U S A 2001 May 8;98(10):5515-20
3. Melanoma-inhibitory activity protein concentrations in blood of melanoma patients treated with immunotherapy.
Oncol Res 1999;11(1):55-61

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中華民國95年06月06日更新