Acrp30 Human, HEK
Adiponectin Human Recombinant, HEK

Formulation:
Sterile filtered and lyophilized from 0.5 mg/ml in PBS buffer.

Solubility:
Add 0.2 ml of deionized water and let the lyophilized pellet of Acrp30 Human dissolve completely.

Stability:
For long term, store lyophilized Acrp30 Human at -20°C. Aliquot the product after reconstitution to avoid repeated freezing/thawing cycles. Reconstituted Acrp30 Human can be stored at 4°C for a limited period of time; it does not show any change after two weeks at 4°C.
The lyophilized Acrp30 Human remains stable for 24 months when stored at -20°C.

Purity:
Acrp30 Human purity is greater than 98% as determined by HPLC and SDS PAGE.

Activity in vitro:
In vitro gluconeogenesis assay in primary hepatocytes was performed, showing the Adiponectin human derived from mammalian cells can inhibit glucose production. The ED50 was ~6 µg/ml.
Effect of Adiponectin human on gluconeogenesis in primary rat hepatocytes.
Primary rat hepatocytes were isolated and maintained as described by Wang et al., 2002. The cells were then treated with 50 pM insulin plus different concentration of Adiponectin Human purified from HEK293 cells for 48 hours. At the end of treatment, the cells were incubated with a glucose-free medium for 6 hours, and glucose production was measured.
Effect of Acrp-30 Human Recombinant on macrophage scanvager receptor A(SRA) expression in THP1-derived macrophages.
THP-1 cells grown in 6-well dishes were differentiated into macrophages using PMA. After starvation in 0.5% FBS culture medium for 16 hours, the cells were treated with 5 µg/ml human recombinant adiponectin for 2 and 3 days, respectively. Total RNA was purified using Trizol reagents. The mRNA abundance of SRA was quantified using quantitative RT PCR (Celera Gene ID: hCG27149), and normalized against 18S RNA (n=4).

References:
·Xu A, Wang Y, Keshaw H, Xu LY, Lam KS, Cooper GJ. The fat-derived hormone adiponectin alleviates alcoholic and nonalcoholic fatty liver diseases in mice. J Clin Invest. 2003; 112:91-100.

· Pajvani UB, Du X, Combs TP, Berg AH, Rajala MW, Schulthess T, Engel J, Brownlee M, Scherer PE. Structure-function studies of the adipocyte-secreted hormone Acrp30/adiponectin. Implications fpr metabolic regulation and bioactivity. J Biol Chem. 2003; 278:9073-85.

· Wang Y, Xu A, Knight C, Xu LY, Cooper GJ. Hydroxylation and glycosylation of the four conserved lysine residues in the collagenous domain of adiponectin. Potential role in the modulation of its insulin-sensitizing activity. J Biol Chem. 2002; 277:19521-19529.

· Yamauchi T, Kamon J, Minokoshi Y, Ito Y, Waki H, Uchida S, Yamashita S, Noda M, Kita S, Ueki K, Eto K, Akanuma Y, Froguel P, Foufelle F, Ferre P, Carling D, Kimura S, Nagai R, Kahn BB, Kadowaki T. Adiponectin stimulates glucose utilization and fatty-acid oxidation by activating AMPactivated protein kinase. Nat Med. 2002; 8:1288-1295.

· Berg AH, Combs TP, Du X, Brownlee M, Scherer PE. The adipocyte-secreted protein Acrp30 enhances hepatic insulin action. Nat Med. 2001; 7:947-953.

Usage:
ProSpec's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.