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Biotin-SP-conjugated AffiniPure® Fab Fragment Donkey Anti-Goat IgG (H+L)
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Target: Goat
Host: Donkey
Antibody Format: Fab Fragment
Specificity: IgG (H+L)
Conjugate: Biotin-SP (long spacer)
Product Category: Fab Fragment Affinity‑Purified Antibodies
Clonality: Polyclonal
RRID: AB_2340399
Fab fragment antibodies are generated by papain digestion of whole IgG antibodies to remove the entire Fc portion, including the hinge region. These antibodies are monovalent, containing only a single antigen binding site. The molecular weight of Fab fragments is about 50 kDa.
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.
Buffer: 0.01M Sodium Phosphate, 0.25M NaCl, pH 7.6
Stabilizer: 15 mg/ml Bovine Serum Albumin (IgG-Free, Protease-Free)
Preservative: 0.05% Sodium Azide
Suggested Working Concentration or Dilution Range:
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 (long spacer)
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.