Whole IgG antibodies are isolated as intact molecules from antisera by immunoaffinity chromatography. They have an Fc portion and two antigen binding Fab portions joined together by disulfide bonds and therefore they are divalent. The average molecular weight is reported to be about 160 kDa. The whole IgG form of antibodies is suitable for the majority of immunodetection procedures and is the most cost effective.
Store freeze-dried powder at 2-8°C. When ready to use, rehydrate with indicated volume of d. water and centrifuge if not clear. Product is stable for about 6 weeks at 2-8°C as an undiluted liquid. Prepare working dilution fresh each day. For extended storage after rehydration, add an equal volume of glycerol (ACS grade or better) for a final concentration of 50%, and store at -20°C as a liquid. Note: after the addition of glycerol, the concentration of protein and buffer salts is one-half of the original. Alternatively, aliquot and freeze the product at -70°C or below in the absence of glycerol. Avoid repeated freezing and thawing.
Expiration date: one year from date of rehydration. However, the expiration date may be extended if the product is stored according to the recommendation and the test results are acceptable for its intended use.
1:100 - 1:800 for most applications
Dilution factors are presented in the form of a range because the optimal dilution is a function of many factors, such as antigen density, permeability, etc. The actual dilution used must be determined empirically.
Alexa Fluor® 594-conjugated antibodies absorb light maximally around 591 nm and fluoresce with a peak around 614 nm. They are brighter, more photostable, and more hydrophilic than Texas Red conjugates. Alexa Fluor® 594 conjugates are brighter than red-fluorescing conjugates, and they provide more color separation from green-fluorescing dyes than DyLight 549, Cy3, and TRITC conjugates. They are the best choice for immunofluorescence detection in the deep-red region of the visible spectrum.
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