Target: Chicken
Host: Donkey
Antibody Format: F(ab')₂ Fragment
Specificity: IgY (IgG) (H+L)
Minimal Cross Reactivity: Bovine, Goat, Guinea Pig, Syrian Hamster, Horse, Human, Mouse, Rabbit, Rat, Sheep Serum Proteins
Conjugate: Alexa Fluor® 647
Product Category: F(ab')₂ Fragment Affinity-Purified Antibodies
Clonality: Polyclonal
RRID: AB_2340380

Information

Based on immunoelectrophoresis and/or ELISA, the antibody reacts with whole molecule chicken IgY. It also reacts with the light chains of other chicken 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, goat, guinea pig, syrian hamster, horse, human, mouse, rabbit, rat and sheep serum proteins, but it may cross-react with immunoglobulins from other species.

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.

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 pepsin 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:

1:100 - 1:800 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.

Conjugate

Alexa Fluor® 647

Amax: 651 Emax: 667nm

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.

Alexa Fluor® 647 AffiniPure™ F(ab')₂ Fragment Donkey Anti-Chicken IgY (IgG) (H+L)