LIF, or Leukemia Inhibitory Factor, belongs to the IL-6 family of cytokines, and is a variably glycosylated, secreted protein with a mass of 38-67kD. The mature form has 180 amino acids, and contains six conserved cysteines which form three intramolecular disulfide bonds. The protein itself forms a four-helix bundle fold. There are two isoforms of mouse LIF formed by alternative splicing that differ by their localization signals. Mouse LIF receptor has one transmembrane and two soluble forms. Mouse LIF shares 92% and 78% amino acid identity with rat LIF and human LIF, respectively.
LIF is a pleiotropic cytokine involved in cellular development, inflammation, immunology, and neoplasia. It often has contradictory effects within different contexts, either stimulating cell proliferation, or inducing differentiation and apoptosis. LIF was originally named for its observed function to promote differentiation of mouse M1 leukemic myeloid cell line into macrophages. With cultured embryonic stem cells, LIF has been an important laboratory tool to help maintain stem cell pluripotency for cell cultures. It is involved in the mechanisms of embryonic implantation, inflammatory disorders such as arthritis, immunomodulation, carcinogenesis, and neoplasia-associated cachexia.
LIF binds to both LIF Rα and gp130 on the cell surface to form a heterotrimeric complex, which initiates cell signaling through the JAK-STAT pathway. LIF can also activate the MAPK, PI3K, and p53 signaling pathways. Soluble forms of both binding partners do exist in circulation, and potentially serve to further regulate LIF action.
BioLegend’s ELISA MAX™ Mouse LIF ELISA MAX™ Deluxe Set is a sandwich Enzyme-Linked Immunosorbent Assay (ELISA). The got anti-mouse LIF polyclonal capture antibody is first coated on a 96-well plate. Standards and samples are added to the wells and free LIF binds to the immobilized capture antibody. Next, biotinylated goat anti-mouse LIF polyclonal detection antibody is added, producing an antibody-antigen-antibody “sandwich”. Avidin horseradish peroxidase is subsequently added, followed by TMB Substrate Solution, producing a blue color in proportion to the concentration of LIF present in the sample. Finally, the Stop Solution changes the reaction color from blue to yellow, and the microwell absorbance is read at 450 nm with a microplate reader. This ELISA set is specifically designed for the accurate quantitation of mouse LIF from cell culture supernatant, serum, and plasma. It is analytically validated with ready-to-use reagents.
Kit Contents
- Mouse LIF ELISA MAX™ Capture Antibody (200X)
- Mouse LIF ELISA MAX™ Detection Antibody (200X)
- Mouse LIF Standard
- Avidin-HRP (1000X)
- Substrate Solution F
- Coating Buffer A (5X)
- Assay Diluent A (5X)
- Lot-Specific Certificate of Analysis/ELISA MAX™ Deluxe Set Protocol
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