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ReliaTech GmbH Specification/DATA SHEET Recombinant Mouse Leukemia Inhibitory Factor (LIF) 131022BB FOR RESEARCH ONLY! NOT FOR HUMAN USE! Cat.-no: Size: Lot. No.: Country of origin: M30-007 10 µg According to product label Germany Scientific Background Gene: LIF Synonyms: Differentiation-stimulating factor, Melanoma-derived LPL inhibitor, Emfilermin Leukemia Inhibitory Factor also called LIF is a lymphoid factor that promotes long-term maintenance of embryonic stem cells by suppressing spontaneous differentiation. Leukemia Inhibitory Factor has several functions such as cholinergic neuron differentiation, control of stem cell pluripotency, bone & fat metabolism, mitogenesis of factor dependent cell lines & promotion of megakaryocyte production in vivo. Human and mouse LIF exhibit a 78% identity in its amino acid sequence. Human LIF is as active on human cells as is it is on mouse cells, though mouse LIF is about 1000 fold less active on human cells, than human LIF. Recombinant mouse LIF produced in E. coli is a single, non-glycosylated, polypeptide chain containing 180 amino acids and having a molecular mass of 19.86 kDa. Sequence SPLPITPVNATCAIRHPCHGNLMNQIKNQLAQLNGSANALFISYYTAQGEPF PNNVEKLCAPNMTDFPSFHGNGTEKTKLVELYRMVAYLSASLTNITRDQKVL NPTAVSLQVKLNATIDVMRGLLSNVLCRLCNKYRVGHVDVPPVPDHSDKEAF QRKKLGCQLLGTYKQVISVVVQAF Database references Protein RefSeq: NP_032527 Uniprot ID: P09056 mRNA RefSeq: NM_008501 Product Specifications Expressed in E.coli Purity > 98% by SDS-PAGE & silver stain Endotoxin level Buffer < 0.1ng per µg (IEU/µg) of rm LIF silver stain 0.5X PBS Stabilizer None Formulation lyophilized Length (aa): 180 MW: 19.86 kDa Result by Nterminal sequencing SPLPIT Stability: The lyophilized LIF, though stable at room temperature, is best stored desiccated below 0°C. Reconstituted LIF should be stored in working aliquots at -20°C. For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA). Reconstitution: Centrifuge vial prior to opening. The lyophilized LIF should be reconstituted in water to a concentration of 200 µg/ml. Store mLIF in a concentration not less than 100 µg/ml. This solution can be diluted into other buffered solutions or stored at -20 °C for future use. AVOID REPEATED FREEZE AND THAW CYCLES! References 1. Hirai H et al, Biochem J 438:11-23, 2011 2. Mathieu ME et al, Stem Cell Rev. 8:1-15, 2012 3. Metcalfe SM, Genes Immun 12(3):157-68, 2011 4. Broholm C and Pedersen BK, Exerc Immunol Rev 16:77-85, 2010 5. Aghajanova L, Curr Opin Obstet Gynecol 22(3):213-9, 2010 6. Paiva P et al, Cytokine Growth Factor Rev 20(4):319-28, 2009 Biological Activity: The ED50 as determined by the LIF-induced inhibition of M1 cell proliferation is in the range of 0.1 - 0.5 ng/ml. Note: The sale and/or commercial use of recombinant human LIF is prohibited in the United States of America (USA). ReliaTech GmbH Phone: +49 (0)5331 8586 987 Fax: +49 (0)5331 8586 989 E-mail: [email protected] web: www.reliatech.de Location: Lindener Str. 15, 38300 Wolfenbüttel, Germany 1 ReliaTech GmbH Specification/DATA SHEET Recombinant Mouse Leukemia Inhibitory Factor (LIF) LI F Handling/Applications m M 70 50 40 30 25 20 - 15 - (SDS-PAGE 15%, red.) Figure 1. SDS-PAGE analysis of recombinant mouse LIF. Sample was loaded in 15% SDS-polyacrylamide gel under reducing conditions and stained with Coomassie blue. 1.0 Absorbance (562 / 630 nm) 0.8 0.6 0.4 0.2 0.0 0.001 0.01 0.1 1 10 mLIF [ng/ml] Figure 2. Inhibition of the proliferation of M1 cells by increasing amounts of recombinant mouse LIF. ReliaTech GmbH Phone: +49 (0)5331 8586 987 Fax: +49 (0)5331 8586 989 E-mail: [email protected] web: www.reliatech.de Location: Lindener Str. 15, 38300 Wolfenbüttel, Germany 2