rhodamine conjugated ricinus communis agglutinin i  (Vector Laboratories)

Journal: BMC Neuroscience

Article Title: Biochemical enrichment and biophysical characterization of a taste receptor for L-arginine from the catfish, Ictalurus puntatus

doi: 10.1186/1471-2202-5-25

Figure Lengend Snippet: Lectin labelling of solubilized barbel epithelial proteins SDS-PAGE (4–20%) of 10 μg of solubilized barbel homogenate stained with silver ( Lane "Sp" ) or probed with PHA-E lectin ( Lane "PHA" ) or RCA-I lectin ( Lane "RCA" ) using lectins at 10 μg/ml with ABC detection. Both lectins label a band at 82 – 84 kDa and lightly label at least two other bands, one near 88 kDa, the other near 120 kDa.

Article Snippet: Agarose-bound lectins, Ricinus communis agglutinin I (RCA-I), Phaseolus vulgaris Erythroagglutinin (PHA-E), in their biotinylated forms, the ABC kits, and rhodamine-conjugated RCA-I were purchased from Vector Lab (Burlingame, CA).

Techniques: SDS Page, Staining

Journal: BMC Neuroscience

Article Title: Biochemical enrichment and biophysical characterization of a taste receptor for L-arginine from the catfish, Ictalurus puntatus

doi: 10.1186/1471-2202-5-25

Figure Lengend Snippet: RCA-I lectin histochemistry on barbel of catfish, I. punctatus (albino). Barbels were fixed in 4% PFA, PBS, cryostat sectioned at 10 microns and histochemically probed with conjugated RCA-I at 1/200 dilution of the manufacturer's stock. (A). Surface labelling by RCA-I shows preferential recognition of binding sites primarily at the apical endings of taste buds. (B). Labelling by RCA-I of the apical region of two taste buds. ( C ). Labelling by RCA-I of horizontal section through the barbel showing reactive taste buds lining the epithelium.

Article Snippet: Agarose-bound lectins, Ricinus communis agglutinin I (RCA-I), Phaseolus vulgaris Erythroagglutinin (PHA-E), in their biotinylated forms, the ABC kits, and rhodamine-conjugated RCA-I were purchased from Vector Lab (Burlingame, CA).

Techniques: Binding Assay

Journal: BMC Neuroscience

Article Title: Biochemical enrichment and biophysical characterization of a taste receptor for L-arginine from the catfish, Ictalurus puntatus

doi: 10.1186/1471-2202-5-25

Figure Lengend Snippet: The unitary current/voltage (I-V) relationship of channels formed by protein of each enrichment step. The I-V relationships were obtained using L-Arg – active protein from the RCA-I lectin column (o) from the first peak fractions off the Sephacryl S-300 HR column (■) and from the pH 9 elution of the ion-exchange column (▲). Measurements were made under symmetrical conditions of 100 mM NaCl, 1 mM CaCl 2 and 5 mM MOPS (pH = 7.2). Data points indicate the Mean ± S.D. The data sets are well fit by a linear regression ( r = 0.99 solid and dotted lines) with slopes of 58, 67, and 73 pS respectively. Bilayer was DOPS:DOPE, 1:1.

Article Snippet: Agarose-bound lectins, Ricinus communis agglutinin I (RCA-I), Phaseolus vulgaris Erythroagglutinin (PHA-E), in their biotinylated forms, the ABC kits, and rhodamine-conjugated RCA-I were purchased from Vector Lab (Burlingame, CA).

Techniques:

Journal: BMC Neuroscience

Article Title: Biochemical enrichment and biophysical characterization of a taste receptor for L-arginine from the catfish, Ictalurus puntatus

doi: 10.1186/1471-2202-5-25

Figure Lengend Snippet: Lectin labelling of solubilized barbel epithelial proteins SDS-PAGE (4–20%) of 10 μg of solubilized barbel homogenate stained with silver ( Lane "Sp" ) or probed with PHA-E lectin ( Lane "PHA" ) or RCA-I lectin ( Lane "RCA" ) using lectins at 10 μg/ml with ABC detection. Both lectins label a band at 82 – 84 kDa and lightly label at least two other bands, one near 88 kDa, the other near 120 kDa.

Article Snippet: Rhodamine-conjugated RCA-I (Vector Labs) was used to estimate the specificity of lectin interaction with glycoproteins of catfish barbel.

Techniques: SDS Page, Staining

Journal: BMC Neuroscience

Article Title: Biochemical enrichment and biophysical characterization of a taste receptor for L-arginine from the catfish, Ictalurus puntatus

doi: 10.1186/1471-2202-5-25

Figure Lengend Snippet: RCA-I lectin histochemistry on barbel of catfish, I. punctatus (albino). Barbels were fixed in 4% PFA, PBS, cryostat sectioned at 10 microns and histochemically probed with conjugated RCA-I at 1/200 dilution of the manufacturer's stock. (A). Surface labelling by RCA-I shows preferential recognition of binding sites primarily at the apical endings of taste buds. (B). Labelling by RCA-I of the apical region of two taste buds. ( C ). Labelling by RCA-I of horizontal section through the barbel showing reactive taste buds lining the epithelium.

Article Snippet: Rhodamine-conjugated RCA-I (Vector Labs) was used to estimate the specificity of lectin interaction with glycoproteins of catfish barbel.

Techniques: Binding Assay

Journal: BMC Neuroscience

Article Title: Biochemical enrichment and biophysical characterization of a taste receptor for L-arginine from the catfish, Ictalurus puntatus

doi: 10.1186/1471-2202-5-25

Figure Lengend Snippet: The unitary current/voltage (I-V) relationship of channels formed by protein of each enrichment step. The I-V relationships were obtained using L-Arg – active protein from the RCA-I lectin column (o) from the first peak fractions off the Sephacryl S-300 HR column (■) and from the pH 9 elution of the ion-exchange column (▲). Measurements were made under symmetrical conditions of 100 mM NaCl, 1 mM CaCl 2 and 5 mM MOPS (pH = 7.2). Data points indicate the Mean ± S.D. The data sets are well fit by a linear regression ( r = 0.99 solid and dotted lines) with slopes of 58, 67, and 73 pS respectively. Bilayer was DOPS:DOPE, 1:1.

Article Snippet: Rhodamine-conjugated RCA-I (Vector Labs) was used to estimate the specificity of lectin interaction with glycoproteins of catfish barbel.

Techniques: