This Mouse Lactoferrin/LTF overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Lactoferrin/LTF protein (Cat: 50750-M08H) from the overexpression lysate was verified.
A DNA sequence encoding the mouse LTF (AAH06904.1) (Met 1-Gln 707) was expressed, with a C-terminal polyhistidine tag.
The secreted recombinant mouse LTF comprises 699 amino acids and has a calculated molecular mass of 77.2 kDa. The apparent molecular mass of the recombinant protein is approximately 70 kDa in SDS-PAGE under reducing conditions.
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.
Lactotransferrin, also known as Lactoferrin, Talalactoferrin, and LTF, is a secreted protein that belongs to the transferrin family. Transferrins are iron binding transport proteins that can bind two Fe3+ ions in association with the binding of an anion, usually bicarbonate. Lactotransferrin has antimicrobial activity which depends on the extracellular cation concentration. Lactoferroxins A, B, and C have opioid antagonist activity. Lactoferrin A shows a preference for mu-receptors, while lactoferricin B and lactoferricin C have somewhat higher degrees of preference for kappa-receptors than for mu-receptors. Lactoferrin / LTF is a globular glycoprotein that is widely represented in various secretory fluids, such as milk, saliva, tears, and nasal secretions. Lactoferrin / LTF is also present in secondary granules of PMN and is secreted by some acinar cells. Lactoferrin / LTF can be purified from milk or produced recombinantly. Human colostrum has the highest concentration, followed by human milk, then cow milk. Lactoferrin / LTF is one of the components of the immune system of the body; it has antimicrobial activity (bacteriocide, fungicide) and is part of the innate defense, mainly at mucose. In particular, lactoferrin provides an antibacterial activity to human infants. Lactoferrin interacts with DNA and RNA, polysaccharides and heparin, and shows some of its biological functions in complexes with these ligands.