YARIV REAGENTS
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YARIV REAGENTS
FOR DETECTION AND QUANTITATION OF
ARABINOGALACTAN-PROTEINS
FOR RESEARCH PURPOSES ONLY
Cat. Nos. 100-2, 100-3, 100-4, 100-5, 100-6, 100-8
R = b-D-glucosyl (Cat. No. 100-2), a-D-galactosyl (Cat. No. 100-3), a-D-mannosyl (Cat. No. 100-5),
b-D-mannosyl residue (Cat. No. 100-6) or b-D-galactosyl (Cat. No. 100-8)
APPLICATIONS
The b-glucosyl Yariv reagent can be used as;
· A histochemical reagent to detect AGP's intissue sections (Anderson et al., 1977;Gleeson & Clarke, 1979).
· To detect and quantify the amounts ofAGP's in a tissue extract (Van Holst &Clarke, 1985).
· To separate AGP's according to theircharge by crossed electrophoresis (VanHolst & Clarke, 1986).
CHEMICAL PROPERTIES
The b-glucosyl Yariv reagent [1,3,5-tris (4-b-Dglycopyranosyloxyphenylazo)-
2,4,6-trihydroxybenzene]are red dyes which specifically bindto and precipitate a class of plant proteolysis,the AGP's (Yariv et al., 1967; Jermyn & Yeow,1975; Anderson et al., 1977; Clarke et al.,1979; Fincher et al., 1983).
METHOD OF USE
As a histochemical reagent:
Dissolve 2mg of b-glucosyl Yariv reagent in1mL of 0.15M NaC1. Apply to tissue sectionfor 1h at room temperature and examine bybright field microscopy. The AGP's precipitatein the Yariv reagent to give a red stain. Theexact colour varies from brown-red to brightred depending on the tissue. As AGP's areusually water-soluble, they may be lost duringthe procedures for tissue embedding; it istherefore advisable to start with hand orcryostat sections. As a control the a-Gal Yarivreagent is used.
For quantification:
Prepare a solution containing 1% w/v agarose,0.15M NaC1, 0.02% w/v NaN3 and 10?g/mL
?-glucosyl Yariv reagent. Heat to boiling, pour3.5mL aliquots onto preheated glass plates(5x7cm); cool. Punch wells in gel formed.Place test solutions in wells; construct
standard curve using reference AGP (e.g. gumarabic) according to Van Holst & Clarke
(1985).
For crossed electrophoresis:
Prepare a solution containing 1%w/v agarosein 0.025M Tris, 0.2M glycine, 0.02%w/v NaN3
and 30mg/mL ?-glucosyl Yariv reagent. Heatto dissolve agarose; use in crossedelectrophoresis experiments according to VanHolst & Clarke (1986).
All solutions are stable for at least a year atroom temperature.
In some applications the b-D-Mannosyl Yarivreagent can replace b-D-Glucosyl Yariv
reagent. Similarly a-D-Mannosyl Yariv reagentcan replace a-D-Galactosyl Yariv reagent.
YARIV REAGENTS