Human Frataxin quantification in cell lysates
Drug discovery and bioprocess often require accurate quantification technologies to improve the productivity workflow. Current methods such as ELISA do not properly fit with such objectives as they require multiple sample-handling and protocol steps. The human Fc kit enables a rapid quantification of human IgG, and any proteins containing a human Fc region. The assay is compatible with natural IgG as well as recombinant antibodies.
The human Fc kit is a competitive immunoassay for the measurement of hIgGs or any hFc-tagged proteins in a range of biological materials. The assay uses an Eu3+-Cryptate conjugated antibody specific to hFc and d2 coupled hIgGs. hFc provided by the sample competes with d2-labeled human IgG for binding to anti-human Fc Eu Cryptate.
|Detection limit:||9 ng/mL|
|Dynamic range:||5.5 to 4,000 ng/mL|
|Incubation time:||2h30 to ON|
|EC50:||216 - 296 ng/mL|
Figure 1: Various concentrations of four different human Fc chimeras were tested using the Total human IgG assay, following standard assay protocols.
Figure 2 demonstrates the compatibility of the HTRF human Fc assay with different media, even in the presence of 10% FCS. No impact on assay performance was seen. Unlike other immunoassays involving streptavidin-biotin detection, this assay is insensitive to the presence of culture medium, whose quenching effects can be compensated for by our patented 665/620 ratio.
Figure 2: IgG standard curve was drawn up using IgG in phosphate buffer, culture media and RPMI containing 10% FCS+/- 4.4% PSGHT (Penicillin, Streptamycin, Glutamine, Hypoxanthine, Thymidine).
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