Recombinant Human MIF protein, His-tagged

• Cat.No.: MIF-23H
• Product Overview: Recombinant Human MIF protein, fused to His-tag at C-terminus, was expressed in Escherichia coli.

Specification

• Species: Human
• Source: E.coli
• Tag: His
• Protein Length: 123
• Description: Migration Inhibitory Factor (MIF) is a secreted protein without a cleavable signal sequence and is secreted via a specialized, non-classical pathway. It is secreted by macrophages upon stimulation by bacterial lipopolysaccharide (LPS), or by M.tuberculosis antigens. MIF consists of two α-helices and six β-strands, four of which form a β-sheet. The two remaining β-strands interact with other MIF molecules, creating a trimer. Structure-function studies suggest MIF is bifunctional with segregated topology. The N- and C-termini mediate enzyme activity (in theory). Phenylpyruvate tautomerase activity (enol-to-keto) has been demonstrated and is dependent upon Pro at position 1. Amino acids 50-65(a.a.) have also been suggested to contain thiol-protein oxidoreductase activity. MIF has proinflammatory cytokine activity centered around (a.a.) 49 - 65. On fibroblasts, MIF induces, IL-1, IL-8 and MMP expression; on macrophages, MIF stimulates NO production and TNF-α release folllowing IFN-γ activation. MIF apparently acts through CD74 and CD44, likely in some form of trimeric interaction. Human MIF is active on mouse cells. Human MIF is 90%, 94%, 95%, and 90% aa identical to mouse, bovine, porcine and rat MIF, respectively.
• Form: Lyophilized from a 0.2μm filtered concentrated solution in PBS, pH 7.4.
• Bio-activity: Fully biologically active when compared to standard. The specific activity is determined by binding rhCD74 in a functional ELISA.
• Molecular Mass: Approximately 13.5 kDa, a single non-glycosylated polypeptide chain containing 117 amino acids, with 6 × His at the C-terminus.
• AA Sequence: MPMFIVNTNVPRASVPDGFLSELTQQLAQATGKPPQYIAVHVVPDQLMAFGGSSEPCALCSLHSIGKIGGAQNRSYSKLLCGLLAERLRISPDRVYINYYDMNAANVGWNNSTFALEHHHHHH
• Endotoxin: Less than 1 EU/µg of rHuMIF, His as determined by LAL method.
• Purity: >95% by SDS-PAGE and HPLC analysis.
• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 12 months from date of receipt, -20 to -70 centigrade as supplied. 1 month, 2 to 8 centigrade under sterile conditions after reconstitution. 3 months, -20 to -70 centigrade under sterile conditions after reconstitution.
• Reconstitution: We recommend that this vial be briefly centrifuged prior to opening to bring the contents to the bottom. Reconstitute in sterile distilled water or aqueous buffer containing 0.1 % BSA to a concentration of 0.1-1.0 mg/mL. Stock solutions should be apportioned into working aliquots and stored at ≤-20 centigrade. Further dilutions should be made in appropriate buffered solutions.

Gene Information

• Gene Name: MIF
• Official Symbol: MIF
• Synonyms: MIF; macrophage migration inhibitory factor (glycosylation-inhibiting factor); GLIF; macrophage migration inhibitory factor; GIF; L-dopachrome isomerase; L-dopachrome tautomerase; phenylpyruvate tautomerase; MMIF
• Gene ID: 4282
• mRNA Refseq: NM_002415
• Protein Refseq: NP_002406
• MIM: 153620
• UniProt ID: P14174

Citation

First evidence that oligopyridines, α-helix foldamers, inhibit Mcl-1 and sensitize ovarian carcinoma cells to Bcl-xL-targeting strategies.

Journal: Journal of medicinal chemistry    PubMed ID: 25585174    Data: 2015/5/7

Authors: Céline Gloaguen, Anne Sophie Voisin-Chiret, Laurent Poulain

Article Snippet:Capture of His-Bcl-xL (same condition as His-Mcl-1) was performed on channel Fc1 that was used as a reference surface for nonspecific binding measurements.Capture of His-Bcl-xL (same condition as His-Mcl-1) was performed on channel Fc1 that was used as a reference surface for nonspecific binding measurements.. Recombinant human Mcl-1 protein, fused to His-tag, and recombinant human Bcl-xL, His-tagged (DNA sequence encoding the human BCL2L1 isoform 1 (Met 1?Arg 212) fused with a polyhistidine tag at the C-terminus) were purchased from Creative BioMart (45?16 Ramsey Road, Shirley, NY 11967, USA). . A lowmass weight-single-cycle kinetics (LMW-SCK) analysis to determine association, dissociation, and affinity constants (ka, kd, and Kd respectively) was carried out by injecting different protein concentrations.A lowmass weight-single-cycle kinetics (LMW-SCK) analysis to determine association, dissociation, and affinity constants (ka, kd, and Kd respectively) was carried out by injecting different protein concentrations.