A Method for Producing Specific Antisera With Small Doses of Immunogen

Vaitukaitis, Judith L.; Robbins, John B.; Nieschlag, Eberhard; Ross, Griff T.

doi:10.1210/jcem-33-6-988

Cited by 1,346 publications

(260 citation statements)

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“…Mouse anti-T. gondii serum was obtained by infecting CBA mice with the 76 K T. gondii strain and collecting the serum 2 months later. Polyclonal antibodies against MIC3 were obtained from a rabbit subcutaneously immunized with immunoaffinity purified MIC3 according to Vaitukaitis et al (1971). Rabbit anti-ROP1 antiserum had been obtained against a recombinant ROP1-b-galactosidase fusion protein (Saffer et al, 1992) Immunopurification of MIC3 and N-terminal sequence determination Immunosorbent production and immunosorption procedure were performed as described previously (Achbarou et al, 1991).…”

Section: Antibodiesmentioning

confidence: 99%

The microneme protein MIC3 of Toxoplasma gondii is a secretory adhesin that binds to both the surface of the host cells and the surface of the parasite

Lebrun²,

et al. 2000

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SummaryAssay of the adhesion of cultured cells on Toxoplasma gondii tachyzoite protein Western blots identified a major adhesive protein, that migrated at 90 kDa in non-reducing gels. This band comigrated with the previously described microneme protein MIC3. Cellular binding on Western blots was abolished by MIC3-specific monoclonal and polyclonal antibodies. The MIC3 protein affinity purified from tachyzoite lysates bound to the surface of putative host cells. In addition, T. gondii tachyzoites also bound to immobilized MIC3. Immunofluorescence analysis of T. gondii tachyzoite invasion showed that MIC3 was exocytosed and relocalized to the surface of the parasite during invasion. The cDNA encoding MIC3 and the corresponding gene have been cloned, allowing the determination of the complete coding sequence. The MIC3 sequence has been confirmed by affinity purification of the native protein and N-terminal sequencing. The deduced protein sequence contains five partially overlapping EGF-like domains and a chitin binding-like domain, which can be involved in protein±protein or protein± carbohydrate interactions. Taken together, these results suggest that MIC3 is a new microneme adhesin of T. gondii.

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Section: Antibodiesmentioning

confidence: 99%

The microneme protein MIC3 of Toxoplasma gondii is a secretory adhesin that binds to both the surface of the host cells and the surface of the parasite

Lebrun²,

et al. 2000

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“…Antiserum against CCK-4 was prepared by conjugation to thyroglobulin by the carbodiimide method (Vaitukaitis et al 1971). Briefly, 25 mg of thyroglobulin in 0.5 ml of distilled water (dH 2 O) was added to 5 mg of CCK-4 dissolved in 0.5 ml of distilled water and adjusted to pH 5.5.…”

Section: Biochemical Analysesmentioning

confidence: 99%

Sensitivity to CCK-4 in Women with and without Premenstrual Dysphoric Disorder (PMDD) During Their Follicular and Luteal Phases

Mellédo

Merani

Koszycki

et al. 1999

Neuropsychopharmacology

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“…Two rabbits were immunised by an intradermic injection of 100 µg of equine trypsin (emulsified in 0.05 M phosphate buffer, 0.15 M NaCl, pH 7.4 supplemented with complete Freund adjuvant) followed by booster injections of 50 µg every 15 days [7,48]. Blood samples were collected 12 days after each booster injection.…”

Section: Immunisationmentioning

confidence: 99%

Equine trypsin: purification and development of a radio-immunoassay

et al. 2003

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-Shock is accompanied by generalised splanchnic hypoperfusion, and splanchnic organs like the pancreas can be damaged, as shown in animal experimental models and in humans, by the presence of high plasma concentrations of trypsin and other pancreatic enzymes. In order to design a radioimmunoassay technique (RIA) for the measurement of equine trypsin-like immunoreactivity (TLI) in biological fluids, trypsin was purified (with purity 96 %) from the equine pancreas by extraction in an acid medium, ammonium sulfate precipitations, gel filtration chromatography and, after activation of trypsinogen into trypsin, affinity chromatography. Gel polyacrylamide electrophoresis showed a monomeric enzyme with a molecular weight of 27 kDa. The purified equine trypsin served for the immunisation of rabbits in order to obtain a specific antiserum, and the labelled antigen was prepared by iodination of equine trypsin with 125 I. The RIA was based on the binding of the antigen to the antibody followed by the separation of the antigen-antibody complex by immunoprecipitation in the presence of sheep anti-rabbit gammaglobulins and the assay of the radioactivity in the precipitate. The RIA showed good sensitivity, specificity, precision, accuracy and reproducibility. The reference mean value of TLI in the plasma of healthy horses (n = 20) was 30.01 ± 6.84 ng/mL (upper confidence limit 50.52 ng/mL; p < 0.01). Three horses with non strangulating intestinal obstruction without shock showed TLI values within normal limits whereas 5 of 7 horses with strangulation obstruction showed TLI levels above the upper confidence limit. Further studies using the RIA and the enzymatic assay should be performed in order to confirm the role of the pancreas in equine intestinal obstruction. equine trypsin / purification / radio-immunoassay / reference range / intestinal obstruction ³

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A Method for Producing Specific Antisera With Small Doses of Immunogen

Cited by 1,346 publications

References 0 publications

The microneme protein MIC3 of Toxoplasma gondii is a secretory adhesin that binds to both the surface of the host cells and the surface of the parasite

The microneme protein MIC3 of Toxoplasma gondii is a secretory adhesin that binds to both the surface of the host cells and the surface of the parasite

Sensitivity to CCK-4 in Women with and without Premenstrual Dysphoric Disorder (PMDD) During Their Follicular and Luteal Phases

Equine trypsin: purification and development of a radio-immunoassay

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