This Mouse Chemerin/RARRES2 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Chemerin/RARRES2 protein (Cat: 50024-M02H) from the overexpression lysate was verified.
A DNA sequence encoding the mouse Rarres2 (NP_082128.1) (Met1-Ser156) was expressed with the Fc region of human IgG1 at the C-terminus.
The recombinant mouse Rarres2 consists 378 amino acids and predicts a molecular mass of 42.6 kDa.
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Retinoic acid receptor responder protein 2 (RARRES2) is a small secreted protein involved in multiple cancers, including adrenocortical carcinoma (ACC). Serum RARRES2 may be used as a novel prognostic marker for ACC. Retinoic acid receptor responder 2 (RARRES2) is transcriptionally downregulated in multiple cancer types. Previous studies suggested that it can serve as an immune-dependent tumor suppressor by acting as a chemoattractant to recruit anticancer immune cells expressing its receptor, the chemerin chemokine receptor 1 (CMKLR1), to sites of tumor. Mechanistically, RARRES2 overexpression in ACC cells inhibited Wnt/beta-catenin pathway activity by promoting beta-catenin phosphorylation and degradation, it also inhibited the phosphorylation of p38 mitogen-activated protein kinase. Thus RARRES2 is a novel tumor suppressor for ACC, which can function through an immune-independent mechanism.