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because of clinical pessimism or financial constraints. Flow cytometry has been used to diagnose and characterize IMHA in dogs 9–12 and humans. 13–15 This method appears more sensitive than standard agglutination tests for IgG detection. 9–11,13–15 A

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in American Journal of Veterinary Research

valuable proteins, including IgG. Equine plasma collection facilities strive to harvest high-volume, high-quality plasma as frequently as possible without causing sustained protein depletion in donor horses. The purpose of the study reported here was to

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in American Journal of Veterinary Research

prepared and evaluated for agglutination. In the indirect immunofluorescence test, 5 paraformaldehyde-fixed neutrophils were incubated with serum from neutropenic cats and subsequently incubated with fluorescein-labeled anti-cat IgG. Cells were viewed with

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in American Journal of Veterinary Research

dilutions of sera were applied to IFA slides, which were washed and reacted with fluorescein isothiocyanate anti-canine IgG. c Endpoint titers were determined as the last dilution at which brightly stained organisms could be detected with a fluorescence

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in American Journal of Veterinary Research

. Lightning-Link PerCP Conjugation Kit, Innova Biosciences, Cambridge, England. g. Lightning-Link APC Conjugation Kit, Innova Biosciences, Cambridge, England. h. Normal mouse IgG2a K, BD, Franklin Lakes, NJ. i. Rabbit normal IgG, X0903, Dako, Glostrup

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in American Journal of Veterinary Research

IgG by use of reducing SDS-PAGE. h Reactivity of purified anti-cCP IgG and antiserum was verified by precipitation testing against pure cCP, canine serum, and BSA by use of radial double immunodiffusion in agar gel (Ouchterlony technique). i A

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in American Journal of Veterinary Research

% paraformaldehyde, then stained for VE-cadherin with rabbit polyclonal anti-CD144 antibody or for vWF with rabbit polyclonal-anti vWF antibody. Fluorescent-labeled goat anti-rabbit IgG was used as the secondary antibody. Cell nuclei were stained with blue

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in American Journal of Veterinary Research

other studies 3,5,11 but only at 30-day intervals. Those investigators recommended the 30-day interval between plasmapheresis procedures to allow donor horses time to replenish depleted total protein, albumin, and IgG concentrations. Despite the use of

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in American Journal of Veterinary Research

of rabbit anti-canine C3 antiserum, e plates were incubated for another 3 hours at 37°C and then washed with buffered washing solution. Alkaline phosphatase–labeled goat anti-rabbit IgG f (1:1,000 dilution) was added to each well, and plates were

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in American Journal of Veterinary Research

purification was 0.1 mg of anti-cNE IgG/mL of antiserum. A total of 3.2 mg of anti-cNE antibody was purified, and 0.8 mg of anti-cNE antibody was biotinylated for use as the reporter antibody in the ELISA. Use of a 20-fold molar excess of biotin and incubation

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in American Journal of Veterinary Research