Park et al (2000) [140] reported 19 patients with the following symptoms: angioedema (11/19), urticaria (18/19), dyspnea (15/19), Gastrointestinal (10/19)and loss of consciousness (2/19).

Yoshimasu et al. (2000) [244] reported 12 patients with the following symptoms: cutaneous symptoms (9/12), wheezing or asthma (6/12), Gastrointestinal (6/12) and one each of discomfort of larynx, shock and anaphylaxis.

Schiffner et al. (2001) [502] reported urticaria, nausea and mild laryngeal edema in one patient.

Noma et al (2001) [437] reported a case of fatal exercise induced anaphylaxis.

Tanaka et al. (2002) [610] reported data from 20 patients with positive CAP to buckwheat of whom 9 were food allergic, one reacted with asthma to buckwheat as an aeroallergen and 10 others did not show immediate symptoms on ingestion. Of the food allergic patients, 2/9 showed cutaneous symptoms only and 7/9 generalized symptoms.

Park et al. (2000) [140] extracted defatted buckwheat flour with phosphate buffered saline.
Nagata et al. (2000) [724] and Matsumoto et al. (2004) [942] extracted buckwheat flour with sodium bicarbonate.
Tanaka et al. (2002) [610] extracted buckwheat flour with a sodium chloride/sodium bicarbonate solution.
Park et al. (1997) [143] used the purified trypsin inhibitors BWI-1 and BWI-2b (see biochemical data).

Park et al. (1997) [143] tested sera from 12 patients.

Park et al (2000) [140] tested sera from 19 patients with symptoms and 15 asymptomatic patients with positive SPT.

Nagata et al. (2000) [724] used sera from 9 allergic patients and 3 non-allergic controls.

Yoshimasu et al. (2000) [244] used sera from 12 allergic subjects with positive RAST

Noma et al (2001) [437] tested sera from a case of exercise induced anaphylaxis.

Tanaka et al. (2002) [610] used sera from 20 patients with positive CAP to buckwheat of whom 9 were food allergic and one reacted to buckwheat as an aeroallergen.

Matsumoto et al. (2004) [942] used sera from 14 buckwheat allergic patients (6 used in Nagata et al. 2000 [724]).

Park et al. (1997) [143] found that 7/12 subjects were RAST positive for BWI-1 but that only one of five sera tested had more than 50% inhibition. The BWI-1 family are candidates for or components of the 9 kDa allergen.

Park et al (2000) [140] found that all 19 allergic patients gave positive CAP as did 7/15 asymptomatic patients with positive SPT. The allergic patients' sera contained 0.53 to 53.5 kU/L buckwheat specific IgE (2/19 patients were below 1.0 kU/L).

Tanaka et al. (2002) [610] found that CAP-FEIA gave 1.70-87.2 kU/L for buckwheat in the 10 patients buckwheat allergic patients. Patients without food allergy gave 1.17-18.7 kU/L.

Noma et al. (2001) [437] reported RAST scores of 2 for soybeans, 3 for buckwheat, 2 for rice, and 3 for wheat in their patient.

Matsumoto et al. (2004) [942] used 1D-SDS-PAGE with 10 or 15% gels with reduction of samples.

Park et al (2000) [140] used 1D-SDS-PAGE with 13.5% gels with and without reduction of samples, which were boiled for 5 minutes.

Yoshimasu et al. (2000) [244] used 1D-SDS-PAGE on 15% gels with and without reduction. Samples were boiled for 5 minutes.

Tanaka et al. (2002) [610] used 1D-SDS-PAGE on 4-20% gradient gels with reduction. Samples were boiled for 3 minutes.

Lee et al. (2001) [396] used 1D-SDS-PAGE on 4-20% gradient gels.

Matsumoto et al. (2004) [942] and Nagata et al. (2000) [724] transferred proteins onto a PVDF microporous membrane (Immobilon, Millipore, MA). The membrane was incubated with patients' sera at a dilution of 1: 3, followed by the addition of alkaline phosphatase-conjugated anti-human IgG, IgE, or IgA (1 : 1000) and visualized by reaction with 5-bromo-4-chloro-3-indolyl phosphate (BCIP)/nitro blue tetrazolium.

Park et al (2000) [140] transferred proteins onto a 0.45µm nitocellulose membrane by electroblotting. This was blocked with Tris and 5% (w/v) non-fat milk with Tween 20 and incubated with patients' sera at a dilution of 1:10, washed with Tris buffer with Tween 20 and incubated with goat alkaline phosphatase-conjugated anti-human IgE (1:1000), then visualized by reaction with 5-bromo-4-chloro-3-indolyl phosphate (BCIP)/nitro blue tetrazolium.

Yoshimasu et al. (2000) [244] transferred proteins onto a nitocellulose membrane by electroblotting. This was blocked with Tris/BSA with Tween 20 and incubated with patients' sera at a dilution of 1:10, washed Tris buffered saline with Tween 20 and incubated with alkaline phosphatase-conjugated anti-human IgE or IgG, which was visualized by reaction with 5-bromo-4-chloro-3-indolyl phosphate (BCIP)/nitro blue tetrazolium.

Tanaka et al. (2002) [610] transferred proteins onto a PVDF membrane. This was washed with PBS with Brij 35 and blocked with BSA. The membrane was incubated with patients' sera at a dilution of 1:10, followed by the addition of goat alkaline phosphatase-conjugated anti-human IgE, or IgA (1 :5000) and visualized by reaction with 5-bromo-4-chloro-3-indolyl phosphate (BCIP)/nitro blue tetrazolium.

Lee et al. (2001) [396] transferred proteins onto a nitocellulose membrane by electroblotting. This was blocked with Tris/BSA and incubated with patients' sera at a dilution of 1:20, washed Tris buffered saline with Tween 20 and incubated with alkaline phosphatase-conjugated anti-human IgE, which was visualized by reaction with 5-bromo-4-chloro-3-indolyl phosphate (BCIP)/nitro blue tetrazolium.

Matsumoto et al. (2004) [942] report the 24-kDa protein (BW24KD as the most prominent band, which was recognized equally by IgG, IgA, or IgE. The 10 kDa protein (BW10KD) was more strongly IgE reactive comparted with IgG or IgA, in 57% of allergic individuals.

Park et al (2000) [140] identified by immunoblotting 24kD, 19kD, 16kD and 9kD proteins as major allergens. 30 kDa, 43 kDa and 67 kDa allergens were also found. The asymptomatic subjects also reacted to the 24kD, 16kD and 9kD proteins but only one to the 19 kDa proteins (this band is split).

Nagata et al. (2000) [724] found 73, 70, 62, 58 and 54 kDa IgE binding bands without reduction. The 73, 70, 62 and 58 kDa bands gave 56 and 24 kDa, 52 and 24 kDa, 45 and 24 kDa and 43 and 24 kDa respectively on reduction.

Yoshimasu et al. (2000) [244] found IgE reactive bands between 45-66 kDa and fainter bands at 14 and 18 kDa before reduction. After reduction, 45 kDa, 18 kDa and 14 kDa bands bound IgE strongly. The 6 bands at 45-66 kDa gave a protein band at 24 kDa after reduction which did not bind IgE.

Tanaka et al. (2002) [610] found IgE binding to the 24 kDa protein in 19/20 sera and to the 16 and 19 kDa proteins in 9/10 patients with symptoms. As the 16 kDa protein was resistant to pepsin, they conclude it is associated with anaphylaxis.

Noma et al. (2001) [437] reported 16, 20, 24, and 58 kDa bands.

Lee et al. (2001) [396] report 21 bands from 120 to 4 kDa in food allergic subjects and 8 in asthmatics.

Kondo et al (1993) [92] identified a 24kDa protein as a major allergen in sera tested. The allergen was a heterodimer, with paired components of different molecular weights .

8 patients showed positive responses up to 5g. Two negative subjects were positive to open challenge with boiled buckwheat.

Not described