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Detection of antibody secreting hybridomas with diazobenzyloxymethyl paper: An easy, sensitive and versatile assay
- Source :
- Journal of Immunological Methods. 59:269-275
- Publication Year :
- 1983
- Publisher :
- Elsevier BV, 1983.
-
Abstract
- A sensitive and universal assay for the detection of monoclonal antibodies is described. One microliter of each hybrid cell supernatant is transferred to activated diazobenzyloxymethyl paper and, after coating the uncovered activated sites by incubation with bovine serum albumin, the paper is dipped in a solution of 125 I-labeled antigen. After incubation to allow for fixation of antigen to antibody present in the supernatants, the paper is rinsed and autoradiographed. As many as 96 wells can be tested on one paper and more than 20 papers can be treated in 1 day (∼ 2000 wells). The sensitivity and reliability of the method were tested with monoclonal antibodies against a subunit of scorpion ( Androctonus australis ) hemocyanin. Unlike other immunobinding assays the test depends only on antigen-antibody interaction and not on class or subclass of immunoglobulin. It is suitable for a variety of hybridomas.
- Subjects :
- medicine.drug_class
Androctonus australis
medicine.medical_treatment
Immunology
Methylcellulose
Monoclonal antibody
Subclass
Mice
Antigen
medicine
Animals
Immunology and Allergy
Bovine serum albumin
Antibody-Producing Cells
Incubation
Immunoassay
Mice, Inbred BALB C
Hybridomas
biology
Antibodies, Monoclonal
Hemocyanin
biology.organism_classification
Molecular biology
biology.protein
Rabbits
Antibody
Subjects
Details
- ISSN :
- 00221759
- Volume :
- 59
- Database :
- OpenAIRE
- Journal :
- Journal of Immunological Methods
- Accession number :
- edsair.doi.dedup.....4637fca12cc749373cd499cc44a965a1