Journal: Surgery Open Science

Article Title: Highly efficient multiplex genetic engineering of porcine primary fetal fibroblasts

doi: 10.1016/j.sopen.2020.11.003

Figure Lengend Snippet: Optimization of β4GalNT2. Three sgRNA were designed and tested to determine an optimal cut site for β4GalNT2 transfections. (A) A gene map demonstrates candidate sgRNA templates for cut sites in the β4GalNT2 gene. (B) Gene editing efficiency for various β4GalNT2 cut sites is shown by TIDE analysis. A region in E3 and E2 was each tested 3 times, whereas 1 region in E9 was tested twice. Average efficiency is shown with standard deviation displayed as error. (C-D) Phenotype data for cells stained with DBA-lectin are shown with flow cytometry–generated density plots. (C) A porcine WT control is shown. (D) Phenotype data for cells transfected with E2, E3, and E9 are shown.

Article Snippet: The following labeled lectin conjugates were used for DBA staining: fluorescein-labeled DBA (#FL-1031, Vector Labs) and rhodamine-labeled DBA (#RL-1032, Vector Labs).

Techniques: Transfection, Standard Deviation, Staining, Flow Cytometry, Generated

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: