Journals / Etlik Veteriner Mikrobiyoloji Dergisi / 1995 / Cilt: 8 - Sayı: 1

The preparation of.reagents in Elisa techniques

ELİSA Tekniğinde Kullanılacak Reagentlerin Hazırlanması

Pages
238–253
Publication Date
1995-12-01
DOI
—
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AI Summary & Key Takeaways

AI Insight
⚡ TL;DR(Too Long; Didn't Read)

According to demand, the ELISA reagents will be dispatched to Iaboratory all over the country.

📌 3 Key Takeaways

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Objective & Hypothesis:The ELISA assay is an with antigen-antibody reaction system which uses an antibody coupled enzyme with the substrate in directly proportional to the amount of antibody or antigen present in the sample being tested. lmmunoassays are one of the most powerful of all immunochemical techniques.
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Key Findings & Data:This was the basis for the techniques known ELISA (Enzyme Linked lmmunoSorbent Assay) pioneered by Engvall and colleaques quick and easy, yielding information that would be difficult to determine by the othertechniques . (11 ,23) For this assay, a lot of diluents, buffers (coating, washing, substrate and conjugating buffers) antigens , conjugate , substrate and ext. are needed.
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Conclusion & Impact:According to demand, the ELISA reagents will be dispatched to Iaboratory all over the country.
🔬Design:Research Article
📍Scope:Comprehensive Sample
⚙️Variables:ELISA · Reagentler · Ragıp BAYRAKTAR

Abstract

The ELISA assay is an with antigen-antibody reaction system which uses an antibody coupled enzyme with the substrate in directly proportional to the amount of antibody or antigen present in the sample being tested. lmmunoassays are one of the most powerful of all immunochemical techniques. The next stage in the development of enzyme-immunoassays was the linkage of soluble antigensor antibody to an insoluble solid phase in a way in which the reactivity of the immunological component was retanied . This was the basis for the techniques known ELISA (Enzyme Linked lmmunoSorbent Assay) pioneered by Engvall and colleaques quick and easy, yielding information that would be difficult to determine by the othertechniques . (11 ,23) For this assay, a lot of diluents, buffers (coating, washing, substrate and conjugating buffers) antigens , conjugate , substrate and ext. are needed. In this study; these kind of solutions have been prepared, tested and standardized. According to demand, the ELISA reagents will be dispatched to Iaboratory all over the country.

Özet

Son zamanlarda soluble antijen veya antikorların enzim immuoassay ile solid faz hale getirilip teşhis . edilmesinde büyük gelişmeler kaydedilmiştir . immunoassay, diğer immunokimyasal tekniklerin en önemlilerinden biri olup aynı zamanda diğer tekniklerden daha az komplike, daha kolay ve daha çabuktur. ELiSA olarak bilinen enzim aracılığı ile antijen-antikor ölçülmesi tekniğine dayanan bu test Engvall ve arkadaşları , Van Weemen ile Schuurs tarafından bulunup geliştirilmiştir . (11 ,23) ELiSA yöntemi ile antikorların veya antijenlerin saptanmasında ortam ya antikor yada antijen ile kaplanarak duyarlaştırılır ve üzerine konan antijen veya antikor ile reaksiyona girer, üzerine ilave edilen enzim işaretli antiimmunoglobulin ve substrat ile renk değişimi şeklinde substratın parçalanma oranına göre test sonuçlandırılır ve mikropleytte oluşturulan bu reaksiyonda bir pleyt üzerinde çok sayıda numune i şlenebilir . Biz bu araştırmada bu test için gerekli olan reagentleri, konjugeyt, substrat ve konjugasyon solüsyonları üretimi olarak ele alıp ürettik vede laboratuvarımıza göre test edip standardizasyonunu gerçekleştirdik.

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Cite This Article

BAYRAKTAR, R., BAYRAKTAR, L., DAĞISTAN, M., DEMİRALAY, O. (1995). The preparation of.reagents in Elisa techniques. Etlik Veteriner Mikrobiyoloji Dergisi, Cilt: 8 - Sayı: 1, 238–253. https://ajindex.com/articles/120928

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References (13)

  1. ALLEN, J.C., KUNKEL, H.G. 1963: Production and purification of chicken lmmunoglobulins. Science. 139: 229-241.
  2. ASHORA, P., KROHN, K. 1986: Washing of ELI SA plates with running tap water. Journal of immunological Methods. 88: 141-142.
  3. AVRAMEAS, S., TERNYNCK, T., GUESDON, J.L.1978: Coupling of Enzymes to Antibodies and Antigens. Scand. J.lmmunol. 8: 7-23. BARBARA WiLSON,M., NAKANE, P.K.1978: Recent Developmentsin the periodate method of conjugating Horseradish Peroxidase (HRPO) to antibodies. lmmunofluorescense and related Staining Techniques. Elsevier/NorthHolland Biomedical Press. 1: 215-234.
  4. BENEDICT, A.A., BROWN, J.R., HERST, R.T. 1962: The temporal synthesis and some chromatographic and ultracentrifugal characteristic of chicken antibodies. J.Exper.med. 90: 399-411. BRADFORD. M.M. 1976: A rapid and sensitive method for the q ua ntitation of m icrog ram q u antites of protein atilizing the prin ciple of proteind ye binding. Analytical Biochemistry. 72:248-254.
  5. BOORSMA, D.M. STREEFKERT, J.G. 1978: lmproved method for seperation of peroxidase conjugates. lmmunofluorescense and related staining techniques. Elsevier/North Holland Biomedical press. 225-235
  6. CASE, J.T. and ARDANS, A.A. 1985: Non spesific Reactions in an Enzyme Linked lmmunosorbent Assay caused by binding of lmmunoglobulins in situ to Egg-Propagated lnfectious Bronchitis virus. Avian Dis. 30: 149-153.
  7. DIETLIND, H. HERWART,A. 1986: Evolution of low molecular weight lmmunoglobulins, Dev. and Comp. lmmunology. 10: 379-380 ENGVALL,E., CARLSSON, H.E. 1976: Enzyme lmmunoassay ELiSA and EMIT. Methods in Enzymology. 70: 419-438.
  8. ENGVALL, E., CARLSSON, H.E. 1976:
  9. Enyzme-linked lmmunosorbent Assay, ELiSA. First International symposium on lmmunoenzymatic techniques INSERM Symposium No:2: 135-182. HARLOW,E., LANE,D. 1988: Antibodies. A laboratory manual. Cold Spring Harbor Laboratory. 1: 319-359, 553-592.
  10. HATFIELD POL YTECHNIC, Division of Biological and Environmental Sciences, 28-31 MARCH 1988. Malecular Biology update in USA. 125-142.
  11. HEBERT, G.A., PATRICIA, L.P., PITTMAN,B. 1973: Determination of the optimal Ammonium Sulphate concentration for the fractionation. of Rabbit Sheep, Horse and Goat Antisera. Applied Micro. biology. 25: 26-36
  12. JENSENIUS, J.C., ANDERSEN, 1., HAU, J., CRONE, M. and KOCH, C. 1981: Eggs: Conveniently package0 Antibodies. Methods for purification of YOLK lgG. Journal of lmmunulogical Methods. 46: 63-68
  13. LESLIE, G.A., CLEM, L.W. 1969: Phylogeny of lg