Information Based on antigen-binding assay and/or ELISA, the antibody reacts with the Fc portion of mouse IgG2a but not with the Fab portion of mouse immunoglobulins.No antibody was detected against mouse IgM or againstnon-immunoglobulin serum proteins.The antibody may cross-react with other mouse IgG subclasses or with immunoglobulins from other species.

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.

Usage 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 a combination of papain digestion and immunoaffinity chromatography using antigens coupled to agarose beads.Fc fragments and whole IgG molecules have been removed. 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 sequential labeling applications.

To complex with primary antibody in solution, use 1:1 weight ratio of Fab:primary antibody (3:1 molar ratio). Vortex and incubate for 30 minutes at room temperature prior to use. Titrate complex to optimal dilution for assay.

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.

Conjugate

Rhodamine Red™-X (RRX)

Amax: 570Emax: 590nm

RRX (Rhodamine Red-X) conjugates have a peak of excitation at 570 nm and a peak of emission at 590 nm. Although TRITC has been used traditionally with FITC for double labeling, better color separation is achieved by using RRX or Alexa Fluor® 594. Rhodamine Red-X is particularly useful for 3- and 4-color labeling with DyLight 405, Alexa Fluor® 488, and Alexa Fluor® 647 by using a confocal microscope equipped with a 405 nm laser and a krypton/argon laser. Fluorescence from RRX lies about midway between that of Alexa Fluor® 488 and Alexa Fluor® 647, and it shows little overlap with either dye. The krypton-argon laser emits lines at 488 nm, 568 nm, and 647 nm, which are optimal for exciting Alexa Fluor® 488, RRX, and Alexa Fluor® 647, respectively. By adding a 405 nm laser and a 420 nm emission filter, 4-color labeling is possible using DyLight 405-conjugated secondary antibodies from JIR (Figure 5). The separation between all four dyes is perfect for 4-color labeling, and all four dyes are very bright.

Images & References

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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.

Rhodamine Red-X is a trademark of Molecular Probes, Inc. Jackson ImmunoResearch is licensed by Molecular Probes, Inc. to manufacture and sell conjugates of Rhodamine Red-X reactive dye.

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