Phaseolus vulgaris agglutinin is the name ascribed to a family of lectins, each of which consists of four subunits. There are two different types of subunits. One appears to be involved primarily in red cell agglutination and has been designated the “E” subunit (for erythroagglutinin). The other type is involved in lymphocyte agglutination and mitogenic activity and has been termed the “L” subunit (for leucoagglutinin). These subunits combine to produce five isolectins
One of these isolectins has four “E” subunits and is designated PHA-E. PHA-E possesses strong hemagglutinating activity but is a poor mitogen. PHA-L, with four “L” type subunits, does not agglutinate red cells but is a potent mitogen. The other three isolectins, designated E3L1, E2L2, and E1L3, have erythroagglutinating and mitogenic activities proportional to the number of respective “E” or “L” subunits. We have termed the mixture of the five isolectins PHA (E+L). All of these isolectins are glycoproteins with molecular weights of approximately 125,000 and have isoelectric points between pH 5.2 for PHA-L and pH 6.0 for PHA-E. PHA-L, although a potent mitogen, has a rather narrow range of concentrations for mitogenic stimulation and appears to require calcium and manganese ions for activity. The presence of some “E” subunits in PHA appears to broaden the concentration range of maximum mitogenic stimulation. As a consequence, PHA (E+L) may be preferred for general mitogenic assays. PHA-L has been found to be an excellent, specific marker for use in anterograde neuronal tracing. Fluorescein labeled Phaseolus vulgaris Leucoagglutinin is produced by using the highest quality fluorescein isothiocyanate, our affinity-purified lectin, and special conjugation procedures. Fluorescein labeled Phaseolus vulgaris Leucoagglutinin has an appropriate number of fluorochromes bound which provide the maximum fluorescence and optimum staining characteristics for this particular lectin. This lectin is supplied essentially free of unconjugated fluorochromes and inactive lectin. Accompanying each fluorescent lectin is an analysis data sheet summarizing the results of our quality control tests and providing pertinent information on the product. All of these reagents are supplied as solutions perserved with sodium azide.
Applications:Suitable for use in FLISA. Other applications not tested.
Recommended Dilution:Optimal dilutions to be determined by the researcher.
Elution:100mM acetic acid
Excitation:495nm
Emission:515nm
F/P (molar):5.9
Storage and Stability:May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 6 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
