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Custom Immunology Services
Antibody Characterization Services |
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Several techniques are available to evaluate the quantity, quality, and function of the antibodies produced. These tests include ELISA, SDS PAGE analysis, and isotype determination.
ELISA, Quick, Sensitive, Specific Antibody Screening
In an ELISA, the specific binding between an antigen and antibody is measured using a secondary enzyme labeled antibody and substrate system and is reported as 50% titer. This is the assay of choice for cost-effectively evaluating large numbers of samples and monitoring the progress of an antibody production program. We suggest testing the first bleed of the project to evaluate the effect of antigen on the animal. Information gained from an ELISA at this point will provide an opportunity for you to make an adjustment to your immunization schedule. We will include your pre immunization sera in this assay to assure there is not a pre-immune titer present in your sera. Additional ELISA's, performed later in the project will give you valuable information that can be used to determining which sera samples to include in your research and which bleeds you would like to have purified. ELISA data will be emailed the day it is completed and we will include information to aid in interpretation of data.
Your immunogen may contain components other than your specific antigen. To obtain usable ELISA titer data, coating antigen cannot contain the same non-specific components. For example, if your immunogen is a GST fusion protein, the coating antigen cannot contain the GST or other associated non-specific components. If your immunogen is a peptide conjugated to a carrier protein, your coating antigen can be either the linear peptide or the peptide with an alternative carrier protein as the conjugate.
Electrophoresis / Western blot
The purity of the antibody preparation may be tested (by client request) by polyacrylamide gel electrophoresis. The sample is prepared and loaded into a pre-cast 10% to 12% Tris-HCl denaturing gel. The standards are included with molecular weights ranging from 6.5kD to 200kD. Two bands are observed after staining in Coomassie Blue rep- resenting the heavy chain (55kD) and the light chain (25kD) of the antibody. Each client is provided with a photo of the gel as an indicator of purity.
Isotype Determination
The isotype of murine antibodies (either in supernatants or ascites) can be determined using murine isotyping kits or strips.
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Our Shipping Address
Attn: Immunology Services
465 Swampbridge Road
Denver, PA 17517
tel (800) 345-4114
tel (717) 336-4921
fax (717) 336-3481
AAALAC International Accredited, GLP Studies, OLAW Assured, USDA Research Registered
Document Revision Date: 4/5/07 |
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