YARIV REAGENTSFOR DETECTION AND QUANTITATION OFARA

Biosupplies

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 REAGENTSFOR DETECTION AND QUANTITATION OFARA

YARIV REAGENTSFOR DETECTION AND QUANTITATION OFARA