Although lectins may be isolated from a large array of plants, microorganisms, invertebrates, lower vertebrates and from mammalian cell membranes, Phytohemagglutinin-M (PHA-M) is a compound extracted from the red kidney bean herbaceous angiosperm, (Phaseolus vulgarus). Lectins, some of which are also known as hemagglutinins, are proteins or glycoprotein substances of a non-immune nature, capable of specific recognition of and reversible binding to carbohydrate moieties of complex glyco-conjugates without specifically altering the covalent structure of any of the recognized glycosyl ligands. Lectins may also show a characteristically high specificity for attachment to individual carbohydrate–binding molecules (e.g. sugar moieties) or may bind to a soluble carbohydrate or to a CHO moiety which is part of a glycoprotein or glycolipid complex. They may also cross-link themselves in cell walls or membranes thereby changing the physiology of the membrane per se causing polyagglutination of many different mammalian red blood cell types, altering cell membrane transport systems, altering cell permeability to the ingress of proteins, generally interfering with cellular metabolism, inducing mitosis or other cell biochemical changes to name a few.
Although each batch of PHA-M is tested for performance, the use of Biological Industries PHA-M does not guarantee the successful outcome of any diagnostic testing.
1) Take a vial of Phytohemagglutinin-M (PHA-M) from the specified storage conditions at 2-8°C and read the label.
2) Defrost the PHA-M at room temperature or at 2-8°C. After thawing, each ml will contain 5-10mg of protein.
3) Using aseptic/sterile technique under a laminar-flow culture hood , inoculate approximately 0.5ml of heparinized whole blood into a glass or plastic conveyance with 10ml of medium or in toto,106 viable cells per ml.
4) Incubate the culture at 37°C in a 5% CO2 atmosphere for 72 hours.
5) Recommended Use: two-four (2-4) ml of re-hydrated PHA-M per 100ml of culture medium.
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