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Jackson Immunoresearch

Alexa Fluor® 647 AffiniPure F(ab')2 Fragment Donkey Anti-Guinea Pig IgG (H+L)

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Alexa Fluor® 647 AffiniPure F(ab')2 Fragment Donkey Anti-Guinea Pig IgG (H+L)

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 whole molecule guinea pig IgG. It also reacts with the light chains of other guinea pig immunoglobulins. No antibody was detected against non-immunoglobulin serum proteins. The antibody has been tested by ELISA and/or solid-phase adsorbed to ensure minimal cross-reaction with bovine, chicken, goat, syrian hamster, horse, human, mouse, rabbit, rat and sheep serum proteins, but it may cross-react with immunoglobulins from other species.

Alexa Fluor® 647-conjugated antibodies absorb light maximally around 651 nm and fluoresce maximally around 667 nm. They are brighter than Cy5 and DyLight 650 in aqueous mounting media. Alexa Fluor® 647- and APC-conjugated secondary antibodies are the best choice for flow cytometry when secondary antibodies fluorescing at these wavelengths are desired. Alexa Fluor® 647 conjugates are the best choice of far red-emitting dyes for multiple-labeling detection with a confocal microscope.

A significant advantage of using Alexa Fluor® 647 over lower wavelength-emitting dyes is the low autofluorescence of biological specimens in this region of the spectrum. However, because of its peak emission at 667 nm, Alexa Fluor® 647 cannot be seen well by eye, and it cannot be excited optimally with a mercury lamp. Therefore, Alexa Fluor® 647 is not recommended for use with conventional epifluorescent microscopes. It is most commonly visualized with a confocal microscope equipped with an appropriate laser for excitation and a far-red detector. Alexa Fluor® 647 conjugates are less expensive alternatives to allophycocyanin conjugates for flow cytometry.

Target: Guinea Pig

Host: Donkey

Antibody Format: F(ab')2 Fragment

Specificity: IgG (H+L)

Minimal Cross Reactivity: Bovine, Chicken, Goat, Syrian Hamster, Horse, Human, Mouse, Rabbit, Rat, Sheep Serum Proteins

Conjugate: Alexa Fluor® 647

Clonality: Polyclonal

Physical StateFreeze-dried powder
StorageStore 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 dateOne 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.
PurityThe 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.  
Buffer0.01M Sodium Phosphate, 0.25M NaCl, pH 7.6 
Stabilizer15 mg/ml Bovine Serum Albumin (IgG-Free, Protease-Free) 
Preservative0.05% Sodium Azide 
Suggested Working Concentration or Dilution Range
Histo-/Cyto-Chemistry1:100-1:800 
Flow Cytometry1:100-1:800 
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.

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Product Detail
Thomas No.
C839L46
Mfr. No.
706-606-148
Description
Alexa Fluor® 647 AffiniPure F(ab')2 Fragment Donkey Anti-Guinea Pig IgG (H+L) (min X Bov, Ck, Gt, Sy Hms, Hrs, Hu, Ms, Rb, Rat, Shp Sr Prot), 0.3 mg
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