Tuesday, February 07, 2012

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Archive for the ‘Elisa Assay Questions’ Category

ELISA comes under anyof the following assays? i.e, cell based assay, binding assay,biochemical assay?

Question by allaboutbaby: ELISA comes under anyof the following assays? i.e, cell based assay, binding assay,biochemical assay?

Best answer:

Answer by jonmcn49
ELISA is an acronym for; ” Enzyme linked immunosorbent assay. ” Try binding, as it is bound to its enzyme and then bound by its antibody to an epitope of interest.

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ELISA (Enzyme Linked Immuno Sorbent Assay?

Question by ntinatsepa: ELISA (Enzyme Linked Immuno Sorbent Assay?
blood test: ELISA (Enzyme Linked Immuno Sorbent Assay
what do you know about it?

Best answer:

Answer by hcbiochem
What do you want or need to know about them?

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in ELISA, why the assay need secondary antibody labeled, why no just label the primary antibody?

Question by teck kim: in ELISA, why the assay need secondary antibody labeled, why no just label the primary antibody?

Best answer:

Answer by Isopropyl_Dog
antibodies have two sides: a constant region and a variable region. So the primary antibody has a variable region that specifically binds to the antigen of interest… this primary antibody can be made naturally by the immune system of an organism. If a person is exposed to antigen X, the natural process in his body will create anti-X antibodies. So then we could take a sample of his blood and collect the anti-X antibodies (with affinity chromatography this is easy), purify them and use them in an ELISA to detect antigen X. Unfortunately the natural process does not label the antibodies… thats why we need secondary antibodies.

The secondary antibody specifically binds to the constant region of the primary antibody (all the primary antibodies will have similar constant regions) and has a detectable probe. So you can use the same secondary antibody with lots of different primary antibodies.

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In IFN-g assay, why we collect supernatants of spleen cells after 72 hrs of incubation with antigens?

Question by vijay_hotguy: In IFN-g assay, why we collect supernatants of spleen cells after 72 hrs of incubation with antigens?
For IL-2 bioassay using CTLL cell lines, we use supernatants of splenocytes after 24 hrs of incubation with antigens and for IFN-g Elisa assay it was collected after 72 hrs of incubation. can you explain what might be the reason? Thanks in advance.

Best answer:

Answer by NeuroProf
Yes, IFN-gamma is produced by splenic cells in reaction to infection or allergy. So essentially what you are doing is fooling the cultured cells into thinking there is a major immune response required, and thus they generate IFN-gamma

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