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BiosearchTech: New Blog Article! Molecular Beacons - Lights in the storm http://bit.ly/bgqCm2
Wed, 10 Mar 2010 19:15:07 +0000
| Immunochemicals |
I am planning an ELISA using DNP-BSA to measure the DNP-specific antibody titer in serum. Do you have any information on which conjugation ratio (3 or 31) I should use for this assay?
The higher loaded 31-DNP-BSA should be used for general capture of all anti-DNP-antibodies in an ELISA assay.
The lower loading material could be used to select for high affinity binding antibodies.
When ordering immunochemical conjugates online, I saw that there is a choice for loading. What does this mean? How do I know which loading ratio to choose?
Loading refers to the conjugation ratio, which is the number of hapten groups conjugated to the protein. It is generally accepted that immunization with a lower density hapten-carrier coupled with low doses of antigen preferentially triggers high affinity antibodies. High immunogen doses and high density of hapten on the carrier tend to elicit a higher response since low affinity antibodies are also elicited.
Are Biosearch’s immunochemical products alum precipitated?
Our immunochemical products are not alum precipitated. This must be performed by the researcher. For a protocol on alum precipitation please refer to the following reference:
Seppala, IJ and Makela, O. Adjuvant effect of bacterial LPS and/or alum precipitation in response to polysaccharide and protein antigens. Immunology 1984 Dec, 53(4):827-836
Is it possible to order NP-LPS and NP-BSA with the same NP hapten loading?
This is not possible because LPS is a small molecule polysaccharide (about 3500 Daltons) with very few functional sites for derivitization. Maximum loading is about 2 hapten groups per LPS. On the other hand, BSA protein has a molecular weight of about 60,000 Daltons and has about 60 lysines that can potentially be labeled.
