F(ab')2 fragment antibodies are generated by pepsin digestion of whole IgG antibodies to remove most of the Fc region while leaving some of the hinge region. F(ab')2 fragments have two antigen-binding Fab portions linked together by disulfide bonds and therefore they are divalent. The average molecular weight is about 110 kDa. They are used for specific applications, such as to avoid binding of secondary antibodies to live cells with Fc receptors or to Protein A or Protein G.
Based on immunoelectrophoresis and/or ELISA, the antibody reacts with the Fc portion of mouse IgG heavy chain but not with the Fab portion of mouse immunoglobulins. No antibody was detected against mouse IgM or non-immunoglobulin serum proteins. The antibody has been tested by ELISA and/or solid-phase adsorbed to ensure minimal cross-reaction with human serum proteins, but it may cross-react with immunoglobulins from other species.
Physical State: Freeze-dried solid
Storage: 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.
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.
Purity: The antibody was purified from antisera by a combination of pepsin digestion and immunoaffinity chromatography using antigens coupled to agarose beads. Fc fragments and whole IgG molecules have been removed.
Buffer: 0.01M Tris-HCl, 0.25M NaCl, pH 8.0
Stabilizer: 15 mg/ml Bovine Serum Albumin (IgG-Free, Protease-Free)
Preservative: 0.05% Sodium Azide
Suggested Working Concentration or Dilution Range:
Western Blot:- 1:5,000-1:50,000
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.
Alkaline phosphatase (from calf intestine) conjugates are prepared by a modified method of Avremeas et al., Scand. J. Immunol. 1978. 8 (Supple. 7), 7. Resulting conjugates contain heterogeneous, high molecular weight complexes. They are sensitive reagents for solid-phase immunoassays such as ELISA and Western blotting. Although alkaline phosphatase conjugates are sometimes used for immunohistochemistry, penetration into whole mount tissues may be limited by their large sizes.
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.