Journal: Veterinary Immunology and Immunopathology

Article Title: Generation and characterization of anti-α-enolase single-chain antibodies in chicken

doi: 10.1016/j.vetimm.2010.06.001

Figure Lengend Snippet: Characterization of recombinant α-enolase and polyclonal anti-α-enolase IgY antibodies. Samples in each panel are protein markers (lane M), purified GST (lane 1), purified α-enolase (lane 2), and purified GST-α-enolase fusion protein (lane 3). Purified proteins visualized by Coomassie blue staining (panel A) were blotted onto nitrocellulose paper and probed with anti-GST antibodies (panel B) or sera from 4th-immunized chicken (panel C). The molecular weight of recombinant α-enolase protein is about 48 kD.

Article Snippet: All the proteins were transferred onto nitrocellulose membranes (Amersham Biosciences, UK), which were then blocked with 5% skim milk in TBST for 1 h. Polyclonal goat anti-chicken IgY light chain antibodies (Bethyl Laboratories, Montgomery, TX, USA) were added at 1:3000 dilution and incubated for an additional hour.

Techniques: Recombinant, Purification, Staining, Molecular Weight

Journal: Veterinary Immunology and Immunopathology

Article Title: Generation and characterization of anti-α-enolase single-chain antibodies in chicken

doi: 10.1016/j.vetimm.2010.06.001

Figure Lengend Snippet: Binding activity of scFv antibodies to purified α-enolase analyzed by ELISA. Cellular lysates containing scFv antibodies from randomly selected clones from the 4th panning cycle were examined for their binding to purified α-enolase coated onto the plate wells. Binding activity was detected using the goat anti-chicken light chain antibodies at 1:3000 dilution, followed by HRP-conjugated donkey anti-goat IgG and measured at 450 nm. Two anti-SARS-CoV scFv antibodies (SCoS-S8 and SCoS-L22) were used as negative controls. One additional control experiment was carried out as described without adding primary recombinant scFv antibodies. Polyclonal IgY antibodies from chickens immunized with purified α-enolase were used as a positive control. The ELISA data were represented as means of the duplicated experiments.

Article Snippet: All the proteins were transferred onto nitrocellulose membranes (Amersham Biosciences, UK), which were then blocked with 5% skim milk in TBST for 1 h. Polyclonal goat anti-chicken IgY light chain antibodies (Bethyl Laboratories, Montgomery, TX, USA) were added at 1:3000 dilution and incubated for an additional hour.

Techniques: Binding Assay, Activity Assay, Purification, Enzyme-linked Immunosorbent Assay, Clone Assay, Control, Recombinant, Positive Control

Journal: eLife

Article Title: Optical dopamine monitoring with dLight1 reveals mesolimbic phenotypes in a mouse model of neurofibromatosis type 1

doi: 10.7554/eLife.48983

Figure Lengend Snippet:

Article Snippet: The following antibodies/dilutions were used: rabbit anti-tyrosine hydroxylase (EMD Millipore, AB152, 1:1000; for VAST), monocloncal mouse anti-tyrosine hydroxylase (ImmunoStar, 22941, 1:1000; for TH quantification, somata tracing, and cell counting), polyclonal chicken anti-GFP (Aves, GFP-1020, 1:1000), Alexa Fluor 488-conjugated donkey anti-chicken IgY F(ab’)two fragment (Jackson ImmunoResearch, 703-546-155, 1:1000), Alexa Fluor 647-conjugated donkey anti-rabbit IgG Fab fragment (Jackson ImmunoResearch, 711-606-152, 1:1000), and Alexa Fluor 647-conjugated donkey anti-mouse IgG Fab fragment (Jackson ImmunoResearch, 711-607-003, 1:1000).

Techniques: Recombinant, Plasmid Preparation, Produced, Software, Control, Refractive Index