This product is a mutant protein where all cysteines are replaced with serines. Non-oxidizable chemokine-HMGB1, LPS free, has chemoattractant activity in vitro and in vivo, does not have cytokine-inducing activity and is resistant to inactivation by ROS. This product is produced in E.coli. The product can be used to recruit leukocytes in vivo without inducing cytokine/chemokine production (Venereau et al, 2012). It contains only trace amounts of LPS (<0.4 ng/mg protein), and is tested for the ability y to induce fibroblast migration. LPS free. Measured by its ability to induce migration. Maximal activity in the cell migration assay is obtained at 1 nM.
纯化方法
Purified
纯度
> 95 %
内毒素水平
The purified protein is free from LPS (Cambrex Limulus Amoebocyte Assay QCL-1000, <0.4 ng LPS per mg protein).
HMGB1 is a nuclear protein that is released passively by necrotic cells, retained by apoptotic cells, and secreted actively by inflammatory cells. HMGB1 is essential for life: Hmgb1 knockout mice die shortly after birth.HMGB1 contains three conserved cysteine residues (C23, C45 and C106) and is a redox-sensitive. Chemotaxis-HMGB1 (the form with chemoattractanct activity) is completely reduced, cytokine-HMGB1 contains a disulfide bond between C23 and C45, further oxidation to sulfonates abrogate both activities.In Non-oxidizable chemokine-HMGB1 all cysteines are replaced with serines: these replacements preserve chemoattractant activity in vitro and in vivo, eliminate the cytokine-inducing activity and make the protein resistant to inactivation by ROS. HMGB1 consists of two fairly rigid, L-shaped DNA-binding domains, each referred to as a ‘HMG box’, and an unstructured tail that ends with 30 consecutive negatively charged amino acids. Non-oxidizable chemokine-HMGB1 consists of 215 amino acid residues and has a calculated molecular mass of approximately 24.8 kDa.