Certificate of Analysis & Data Sheet
Resistin, a product of the RSTN gene, is a peptide hormone belonging to the class of cysteine-rich secreted proteins (monomeric peptide contains 11 cysteine residues) referred to as the RELM family, and is also described as ADSF (Adipose Tissue-Specific Secretory Factor) or FIZZ3 (Found in Inflammatory Zone 3). Human resistin is expressed and purified by chromatographic techniques as a 93 amino acid protein without any tag and havin an additional Ala at its N-terminus. It forms a covalent homodimer linked by S-S bond through Cys 7. Resistin may be an important link between obesity and insulin resistance. Resistin, specifically produced and secreted by adipocyte, acts on skeletal muscle myocytes, hepatocytes and adipocytes themselves so that it reduces their sensitivity to insulin. It was suggested that resistin suppressed the ability of insulin to stimulate glucose uptake.
The molecular mass of the purified homodimer is 19576 Da.
Recently it was shown that resistin binds to TLR4 in hypothalamus.
Sterile Filtered White lyophilized (freeze-dried) powder.
Recombinant resistin was lyophilized from a concentrated (1mg/ml) solution containg 0.03% NaHCO3.
It is recommended to reconstitute the lyophilized resistin in sterile 18M-cm H2O not less than 100µg/ml, which can then be further diluted to other buffered aqueous solutions containing carrier proteins.
Lyophilized resistin although stable at room temperature for 3 weeks, should be stored desiccated below -18 C. Upon reconstitution it 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 prevent freeze-thaw cycles.
Greater than 98.0% as determined by:
(a) Analysis by RP-HPLC.
(b) Analysis by reducing and non-reducing SDS-PAGE Silver Stained gel.
(c) Gel filtration chromatography under non denaturative conditions.
Amino Acid Sequence:
The sequence of the first seven N-terminal amino acids was determined to be Ala-Ser-Ser-Lys-Thr-Leu-Cys.
Dimers and Aggregates:
The protein is a covalent homodimer and contains less than 2% aggregates.
The biological activity was evidenced by resistin ability to prevent insulin-stimulated uptake of deoxy glucose in several cell lines at 10 ng/ml concentration. It also induces phosphrylation of Akt in several cell lines.
Less than 0.1 ng/µg (IEU/µg) of human resistin
Protein quantitation was carried out by two independent methods:
1. UV spectroscopy at 280 nm using the absorbency value of 1.26 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 standard solution of BSA as a Reference Standard.
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