Antibody Format: Whole IgG
Specificity: IgG (H+L)
Conjugate: Horseradish Peroxidase
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.
Based on immunoelectrophoresis and/or ELISA, the antibody reacts with whole molecule mouse IgG. It also reacts with the light chains of other mouse immunoglobulins. No antibody was detected against non-immunoglobulin serum proteins. The antibody may cross-react with immunoglobulins from other species.
Physical State: Freeze-dried solid
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.
Purity: The antibody was purified from antisera by immunoaffinity chromatography using antigens coupled to agarose beads.
Buffer: 0.01M Sodium Phosphate, 0.25M NaCl, pH 7.6
Preservative: None (Warning: Use of sodium azide as a preservative will substantially inhibit the enzyme activity of horseradish peroxidase.)
Suggested Working Concentration or Dilution Range:
1:500 - 1:5,000 for immunohisto/cytochemistry
1:5,000 - 1:100,000 for ELISA and Western blotting with chromogenic substrates
1:10,000 - 1:200,000 for Western blotting with ECL substrates
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.
Horseradish peroxidase (HRP) conjugates are prepared by a modified Nakane and Kawaoi procedure (J. Histochem. Cytochem. 1974. 22, 1084). Peroxidase conjugates are commonly used for immunohistochemistry, Western blotting, and ELISA. Affinity-purified anti-horseradish peroxidase and conjugates are available for detection of horseradish peroxidase antigen or for signal amplification of HRP-containing reagents. For immunostaining of mammalian cells, an advantage of using anti-horseradish peroxidase is reduced background, since the antibody does not recognize the endogenous peroxidase-like enzymes found in those cells.