protein

Human Resistin Antagonist

CYT-23

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 antagonist is a monomeric C7A mutant that does not form covalent dimers. It is expressed and purified by chromatographic techniques.

E. coli

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 (BUT NOT FOR in vivo EXPERIMENTS) 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.

The sequence of the first seven N-terminal amino acids was determined to be Ala-Ser-Ser-Lys-Thr-Leu-Ala.

The protein is a covalent monomer and contains less than 2% aggregates.

The biological activity was evidenced by resistin antagonist activity to inhibit resistin-induced Akt phosphorylation in two cell lines. It also reduced the weight (mainly the visceral fat) and normalized GTT and ITT inHFD-fed mice.

Less than 0.1 ng/µg (IEU/µg) of human resistin antagonist

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 computer analysis program of protein sequences (DNAman). 2. Analysis by RP-HPLC, using a standard solution of BSA as a Reference Standard.

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