Certificate of Analysis and Data Sheet

Source:
E.coli

Catalog No.
CYT-433

Introduction:

Adiponectin, also referred to as Acrp30, AdipoQ and GBP-28, is a recently discovered 244 aminoacid protein, the product of the apM1 gene, which is physiologically active and specifically and highly expressed in adipose cells (adipokine). The protein belongs to the soluble defence collagen superfamily; it has a collagen-like domain structurally homologous with collagen VIII and X and complement factor C1q-like globular domain. Adiponectin forms homotrimers, which are the building blocks for higher order complexes found circulating in serum.

Description:

The Human Adiponectin is created as a recombinant protein with N-terminal fusion of HisTag. The Human Adiponectin His-Tagged Fusion Protein, produced in E. coli, is 26.4 kDa protein containing 230 amino acid residues of the human Adiponectin and 12 additional amino acid residues - HisTag (underlined).
MRGSHHHHHH GSGHDQETTT QGPGVLLPLP KGACTGWMAG IPGHPGHNGA PGRDGRDGTP GEKGEKGDPG LIGPKGDIGE TGVPGAEGPR GFPGIQGRKG EPGEGAYVYR SAFSVGLETY VTIPNMPIRF TKIFYNQQNH YDGSTGKFHC NIPGLYYFAY HITVYMKDVK VSLFKKDKAM LFTYDQYQEN NVDQASGSVL LHLEVGDQVW LQVYGEGERN GLYADNDNDS TFTGFLLYHD TN

Specificity:

The amino acid sequence of the recombinant human adiponectin is 100% homologous to the amino acid sequence of the human serum adiponectin.

Purification Method:

Three-step procedure using affinity Ni-NTA chromatography and size exclusion chromatography before and after refolding.

Purity:

>95% (SDS-PAGE analyzed).
SDS-PAGE 12% separation of human adiponectin 5ug / line.

Formulation:

Sterile filtered and lyophilized from 0.5 mg/ml in 0.02M Tris buffer pH7.5, 0.15M NaCl.

Reconstitution:

Add 0.2 ml of 0.1M Acetate buffer pH4 and let the lyophilized pellet dissolve completely. For conversion into higher pH value, we recommend intensive dilution by relevant buffer to a concentration of 10ug/ml. In higher concentrations the solubility of this antigen is limited.

Storage:

Store lyophilized protein at -20蚓. Aliquot the product after reconstitution to avoid repeated freezing/thawing cycles. Reconstituted protein can be stored at 4蚓 for a limited period of time; it does not show any change after two weeks at 4蚓.

Stability/Shelf Life:

The lyophilized protein remains stable until the expiry date when stored at -20蚓.

Quality Control Test:

BCA - to determine content of the protein SDS PAGE - to determine purity of the protein.

Applications:

Western blotting.

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.
?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 AMP-activated 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.

Latest Publications:

1. A longitudinal study of serum adiponectin during normal pregnancy.
BJOG 2006 Jan;113(1):110-3
2. High molecular weight adiponectin correlates with insulin sensitivity in patients with hepatitis C genotype 3, but not genotype 1 infection.
Am J Gastroenterol 2005 Dec;100(12):2717-23
3. Single Nucleotide Polymorphisms in exon 3 of the adiponectin gene in subjects with type 2 diabetes mellitus.
Rocz Akad Med Bialymst 2005;50:148-50
4. Decreased serum adiponectin: an early event in pediatric nonalcoholic fatty liver disease.
J Pediatr 2005 Dec;147(6):835-8
5. Endothelial cell IL-8, a new target for adiponectin: implications in vascular protection.
Circ Res 2005 Dec 9;97(12):1216-9
6. Relationship between plasma adiponectin levels and metabolic risk profiles in Taiwanese children.
Obes Res 2005 Nov;13(11):2014-20