Antibodies immune sera

Williams, D.J. and Behnke, J.M. (1983) Host protective antibodies and serum immunoglobulin isotypes in mice chronically infected or repeatedly immunized with the nematode parasite Nematospiroides dubius. Immunology 48, 37—17. 

Appropriate controls should always be run with any immunocytochemical procedure. Controls may include omitting the primary antibody, substituting pre-immune serum, normal serum, or normal IgG for the primary antibody, adsorbing the primary antibody against the antigen, or immunostaining with an unrelated antibody. 

A simple and fast radioimmunoassay for saxitoxin has been developed using an anti-saxitoxinol antibody. The immune serum required for the immunoassay was raised in rabbits using an immunogen conjugate prepared from saxitoxinol and bovine serum albumin... 

Polyclonal antibodies Passive immunization Serum of immunized animals/ humans... 

To reduce unspecific binding, mix a 200-pl aliquot of the clear supernatant with 2 pi pre-immune serum or unspecific antibody and a further 200 pi aliquot with 50 pi precipitation aid. Rock at 0 °C for 1 h and spin at 1000 x g. Transfer the supernatant into a fresh container and fill it up to 1000 pi with Soln. A. Add 0.5 - 5 pi of the specific antiserum and monoclonal antibody, respectively, and incubate on ice for 1 h. Prepare a second sample containing pre-immune serum instead of antiserum. 

Injected forms of antibodies which have been generated in another body or animal can be isolated, purified, and administered as standard human immune serum globulin (ISG), and ISG plus preparation, or as an animal antiserum or antitoxin. Some serums which are available are those for rabies, snake and insect bites, botulism, and tetanus. Temporary immunity of up to six months to hepatitis can be imparted by one "gamma globulin" shot. More permanent active immunity is available to health care workers. 

The carrier proteins themselves were usually immunogenic, although the antibodies they elicited could be readily adsorbed out of the antisera. In some instances, the contribution of the protein to the specificity of the immune serum raised against the neoglycoprotein was evaluated by attaching the carbohydrate antigen to different carrier-proteins.73... 

The sections are treated with H202 and then incubated in the primary antibody at a dilution of 1 50. This is followed by sequential incubation in the biotinylated antimouse antibody and streptavidin-biotin-labeled complex. DAB is used for 5 min as the chromogen, and the sections are lightly counterstained with hematoxylin. Positive controls involve the use of the tissue known to express the antigen under study. Negative controls involve the replacement of the primary antibody with the diluent alone or with a non-immune serum. 

Prepare three different pools of modified MB-Tosyl magnetic beads (i) with the anti-sulfonamide class-specific antibodies (ii) with the non-specific antibody (with the non-immunized serum) and, (iii) without any antibody, only treated with the corresponding coupling buffer [2],... 

It will be noted in pattern A of Figure 4, that only serum protein was present in the pre-immune serum and antibodies were not eluted by galactose or lactose. Pattern B of the Figure shows that, in addition to the serum proteins, two 280 nm absorbing components were obtained from the anti-S. faecalis serum. One component eluted with galactose and the other eluted with lactose. 

Immune Blood tests Antibody response to immunization (humoral immunity) serum concentrations of immunoglobulin subtypes (humoral immunity)... 

Heneghan, M.A., McCarthy, C.F., Janulaityte, D., Moran, A.P. Relationship of anti-Lewis x and anti-Lewis y antibodies in serum samples from gastric cancer and chronic gastritis patients to Helicobacter py/on -mcdiated autoimmunity. Infect Immun 69 (2001) 4774 1781. 

Polyacrylamide gel electrophoresis is conducted utilizing a published procedure [32], Samples of approximately 50 //g of the antibody solution were subjected to electrophoresis. The buffer solution was of pH 8.3 and consisted of 0.005 M Tris and 0.04 M glycine. Electrophoresis was conducted at a constant current of 2.5 ma/gel for periods of 4 to 6 hrs. The finished gels were stained with 0.02% Coomassie Blue G-250 to reveal the protein components. The results for immune serum and the purified antiglucose antibodies were photographed, Fig. (8A). The non-antibody protein components in the serum have been removed by the affinity method. 

Fig- ( ) Gel electrophoresis patterns of immune serum and affinity purified antibodies [A], Density gradient ultracentrifugation (B), top pattern glucose oxidase, lower pattern mesquite antibody. 

The polysaccharide of Streptococcus pyogenes consists of rhamnose and N-acetyl glucosamine residues. Antibodies have been isolated from S. pyogenes by affinity chromatography by the technique described in an earlier section. The immune serum of S. pyogenes was provided by Dr. McCarty, Rockefeller University. The affinity pattern for the isolation of the antibodies is reproduced in Fig. (12) [45],... 

For polyclonal antibodies, negative reagent controls should be a dilution of immunoglobulin fractions or whole serum of normal/ non-immune serum of the same animal source. Again, the negative reagent control should be applied in the same concentration as the test antibody, and the same diluent should be used. 

Blocking serum or wrong blocking serum used. Block with serum from the host of the secondary or link antibody. Avoid serum that contains auto-immune immunoglobulins. Alternatively, a serum-free protein block, lacking immunoglobulins, may be substituted for the serum block. 115-121... 

IV. Anti-T-cell Immune serum is obtained from animals immunized with human T-lymphocytes. The antibodies bind to and damage T cells and can thus be used to attenuate tissue rejection. 

Fig. 1.—Affinity chromatography elution pattern for anti-rhamnose antibodies in immune serum from Streptococcus mutans. The arrows indicate the point of application of the serum and of 0.5 M L-rhamnose solution. The inset is an agar-diffusion pattern of the antibodies against the antigen. (Reprinted from Carbohydrate Research, Volume 124, J. H. Pazur, M. S. Erikson, M. Tay, and P. Z. Allen, pp. 253-263, copyright 1983, with permission from Elsevier Science Ltd., The Boulevard, Langford Lane, Kidlington 0X5 1GB, UK.)...

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