AMCA AffiniPure Goat Anti-Human IgG (H+L)2

Based on immunoelectrophoresis and/or ELISA, the antibody reacts with whole molecule human IgG. It also reacts with the light chains of other human immunoglobulins. No antibody was detected against non-immunoglobulin serum proteins. The antibody may cross-react with immunoglobulins from other species.

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.

Physical State: Freeze-dried solid

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.

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
Stabilizer: 15 mg/ml Bovine Serum Albumin (IgG-Free, Protease-Free)
Preservative: 0.05% Sodium Azide

Suggested Working Concentration or Dilution Range:

1:50 - 1:200 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.

Coumarin AMCA

Amax: 350 Emax: 450nm

Aminomethylcoumarin Acetate (AMCA) conjugates absorb light maximally around 350 nm and fluoresce maximally around 450 nm. For fluorescence microscopy, AMCA can be excited with a mercury lamp and observed using a UV filter set. Since blue fluorescence is not well detected by the human eye, AMCA-conjugated secondary antibodies should be used only with the most abundant antigens in multiple-labeling experiments. Ways of improving the visibility of AMCA include dark adapting the eyes, using fluorite instead of glass objectives, avoiding mounting media that absorb UV light (such as plastic-based media), and capturing photographic images with blue-sensitive film or CCD cameras. AMCA fades rapidly in conventional epifluorescence and confocal microscopy, and therefore it should be used with mounting media containing an anti-fading agent such as n-propyl gallate.

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