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Biosupplies YARIV REAGENTS

发表时间:2016-08-22  |  点击率:800

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 in
tissue sections (Anderson et al., 1977;
Gleeson & Clarke, 1979).
· To detect and quantify the amounts of
AGP's in a tissue extract (Van Holst &
Clarke, 1985).
· To separate AGP's according to their
charge by crossed electrophoresis (Van
Holst & Clarke, 1986).

CHEMICAL PROPERTIES
The ?-glucosyl Yariv reagent [1,3,5-tris (4-?-Dglycopyranosyloxyphenylazo)-
2,4,6-trihydroxybenzene]
are red dyes which specifically bind
to 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 ?-glucosyl Yariv reagent in
1mL of 0.15M NaC1. Apply to tissue section
for 1h at room temperature and examine by
bright field microscopy. The AGP's precipitate
in the Yariv reagent to give a red stain. The
exact colour varies from brown-red to bright
red depending on the tissue. As AGP's are
usually water-soluble, they may be lost during
the procedures for tissue embedding; it is
therefore advisable to start with hand or
cryostat sections. As a control the ?-Gal Yariv
reagent 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, pour
3.5mL 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. gum
arabic) according to Van Holst & Clarke
(1985).

For crossed electrophoresis:
Prepare a solution containing 1%w/v agarose
in 0.025M Tris, 0.2M glycine, 0.02%w/v NaN3
and 30mg/mL ?-glucosyl Yariv reagent. Heat
to dissolve agarose; use in crossed
electrophoresis experiments according to Van
Holst & Clarke (1986).

All solutions are stable for at least a year at
room temperature.
In some applications the ?-D-Mannosyl Yariv
reagent can replace ?-D-Glucosyl Yariv
reagent. Similarly ?-D-Mannosyl Yariv reagent
can replace ?-D-Galactosyl Yariv reagent.

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