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.
Storage and Rehydration: Store freeze-dried solid at 2-8°C. Rehydrate with the indicated volume of dH2O (see product specification sheet) and centrifuge if not clear. Prepare working dilution on day of use. Product is stable for about 6 weeks at 2-8°C as an undiluted liquid.
Extended Storage after Rehydration: Aliquot and freeze at -70°C or below. Avoid repeated freezing and thawing. Alternatively, add an equal volume of glycerol (ACS grade or better) for a final concentration of 50%, and store at -20°C as a liquid.
Expiration date: one year from date of rehydration. The expiration date may be extended if test results are acceptable for the intended use.
1:20,000 - 1:400,000 for ELISA and Western blotting using enzyme-conjugated streptavidin
1:500 - 1:5,000 for enzyme immunohisto/cytochemistry
1:200 - 1:1,000 for flow cytometry and fluorescence immunohisto/cytochemistry
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.
Biotin-SP is our trade name for biotin with a 6-atom spacer positioned between biotin and the protein to which it is conjugated. When Biotin-SP-conjugated antibodies are used in enzyme immunoassays, there is an increase in sensitivity compared to biotin-conjugated antibodies without the spacer. This is especially notable when Biotin-SP conjugated antibodies are used with alkaline phosphatase-conjugated streptavidin. Apparently, the long spacer extends the biotin moiety away from the antibody surface, making it more accessible to binding sites on streptavidin. Biotinylated antibodies require an additional reagent for visualization. We offer streptavidin and Mouse Anti-Biotin conjugated to fluorophores and enzymes.
Anti-GFP immunohistochemical staining using donkey anti-rabbit IgG secondary antibody (Jackson ImmunoResearch): (A) GFP+ cells were visualized by brown staining in the positive control; (B) The negative control showed no specific immunoreactivity; (C) At 4 weeks post-transplantation, no GFP signal could be detected in the in vivo specimens; (D) PDLCs were GFP positive before implantation.
Sections were pre-incubated in 10% normal donkey serum to inhibit non-specific staining for 60 min at room temperature. Next, sections were incubated with primary antibody (GFP, 1:500, rabbit anti IgG fraction; Molecular Probes) or PBS (negative control) overnight at 4˚C in a humid atmosphere. Subsequently, sections were rinsed with PBS and incubated with a biotin-conjugated donkey anti-rabbit IgG (Jackson ImmunoResearch, West Grove, PA, USA 711-065-152) for 1 h at room temperature. Then, sections were immersed in ABC complex solution in the dark for 45 min at room temperature. The final visualization of reaction products was performed by treatment of 3,3’ diaminobenzidine (DAB) substrate (Envision kit; DakoCytomation, Glostrup, Denmark) for 10 min at room temperature. The nuclei were slightly counter-stained with haematoxylin.
Xiangzhen Yan - Radboud University
This product is for in vitro research use only. It is not a medical device and it is not intended for diagnostic or therapeutic purposes.
Jackson ImmunoResearch Laboratories, Inc. has trademark rights to AffiniPure™.
