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Elizabeth Soberg, University of Washington
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Affinity-purified antibodies are isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. A proprietary elution process is used to dissociate antibodies from the antigen. Unconjugated affinity-purified antibodies are supplied sterile filtered in phosphate buffer without stabilizers or preservatives. Conjugated affinity-purified antibodies are freeze-dried in phosphate buffer with stabilizers and sodium azide, with the exception of horseradish peroxidase conjugates, which do not contain a preservative.
The antibodies are listed alphabetically according to the host species of the antigen (i.e. the primary antibody). For example, if the primary antibody is made in mouse, select "Anti-Mouse". This is the target species in the product filter.
Note: Both anti-Syrian and anti-Armenian hamster secondary antibodies are listed under "Anti-Hamster". It is important to know in which strain of hamster the primary antibody was produced in order to select the proper secondary antibody, since cross-reaction between the strains is not complete.
Selection of the host species for a secondary antibody involves many considerations, including but not limited to:
The following explanations of terms may assist in selecting the most appropriate antibody specificity.
Note: Immunoglobulins from different species share similar structures, with similarities being related to closeness in phylogeny. Antibodies against immunoglobulins from one species may cross-react with a number of other species unless they have been specifically adsorbed against the cross-reacting species. Antibodies that have been adsorbed against other species will contain "(min X...Sr Prot)" in the antibody description.
These antibodies react with both the heavy and light chains of the IgG molecule, i.e. with both the Fc and F(ab')2 / Fab portions of IgG (Figure 1 ). Anti-IgG (H+L) antibodies also react with other immunoglobulin classes (IgM, IgA, IgD, IgE) and subclasses since they all share the same light chains (either kappa or lambda). Anti-IgG (H+L) antibodies have broader epitope recognition than anti-fragment specific antibodies. They are suggested for all general immunodetection procedures.
Anti-IgG, Fc/Fcγ fragment specific
These antibodies react with the Fc portion of the IgG heavy chain. They have been tested by ELISA and/or absorbed against Fab fragments. In some cases, they are additionally tested and/or adsorbed to minimize cross-reactivity to IgM and/ or IgA. In such cases (anti-human, anti-mouse, and anti-rat), they are labeled "Anti-IgG, Fcγ".
Caution: Anti-IgG, Fcγ fragment specific antibodies may not react equally with all monoclonal IgGs. For an anti-mouse IgG, Fcγ fragment specific antibody with balanced reactivity to four subclasses of IgG, select goat anti-mouse IgG (subclasses 1+2a+2b+3), Fcγ fragment specific (min X Hu, Bov, Rb Sr Prot).
Anti-Mouse IgG, Fcγ Subclass specific
These antibodies react with the Fc portion of the heavy chain of individual subclasses of mouse IgG. They have been tested by ELISA and/or adsorbed to minimize cross-reactivity to other subclasses, Fab fragments, IgA, IgM, and a few other species of IgG. Anti-Mouse IgG, Fcγ Subclass specific antibodies are intended for distinguishing between different subclasses of mouse IgG primary antibodies in multiple labeling experiments.
Anti-IgG, F(ab')2 fragment specific
These antibodies react with the F(ab')2 / Fab portion of IgG. They have been tested by ELISA and/or adsorbed against Fc fragments. They are not specific for IgG since they react with light chains, and therefore also react with other immunoglobulin classes (IgA, IgM, IgD, and IgE) and subclasses sharing the same light chains.
(min X ... Sr Prot)
Secondary antibodies against one species may cross-react with other species unless they have been specifically adsorbed against the other species. Antibodies with "(min X ... Sr Prot)" in the description have been tested and/or adsorbed against IgG and/or serum proteins of those species indicated in the parentheses. They are recommended when the presence of immunoglobulins from other species may lead to interfering cross-reactivities. However, caution should be exercised when considering antibodies that have been adsorbed against closely-related species since they have greatly reduced epitope recognition and may recognize some monoclonals poorly. For example, only use anti-mouse IgG adsorbed against rat IgG to detect a mouse primary antibody in rat tissue which contains endogenous rat immunoglobulins, or in a multiple labeling application which includes a rat primary antibody. Use anti-mouse IgG not adsorbed against rat IgG to detect a mouse primary antibody in the absence of rat immunoglobulins. Two other examples of antibodies which have diminished epitope recognition after adsorption with closely-related species are Anti-Rat IgG (min X Mouse Sr Prot) and Anti-Armenian Hamster IgG (min X Mouse, Rat Sr Prot). Refer to "ML (Multiple Labeling)" below for further information.
The following abbreviations are used in the parentheses:
Bov = Bovine
Ck = Chicken
Gt = Goat
GP = Guinea Pig
Sy Hms = Syrian Hamster
Hrs = Horse
Hu = Human
Ms = Mouse
Rb = Rabbit
Shp = Sheep
Sw = Swine
Sr = Serum
Prot = Protein.
ML (multiple labeling)
Some antibodies are designated "ML" to emphasize their usefulness in multiple labeling in addition to single labeling. For further information see Multiple Labeling (ML) Using Labeled Secondary Antibodies.
Most hamster monoclonal antibodies are derived from Armenian hamster spleen cell-mouse myeloma hybridomas. The IgG produced by these hybridomas is Armenian (not Syrian) hamster IgG. Most commercially available polyclonal anti-hamster IgG antibodies have been anti-Syrian hamster IgG, which are not as effective as anti-Armenian hamster IgG in detecting Armenian hamster IgG monoclonal antibodies.
Caution: Anti-Armenian Hamster IgG (H+L) (min X Bov, Hu, Ms, Rb, Rat Sr Prot) may not recognize all Armenian hamster monoclonal antibodies, since it has been adsorbed against closely related species (in bold). Therefore, it is better to use an antibody adsorbed against fewer species, such as Anti-Armenian Hamster IgG (H+L) (min X Bov Sr Prot), except in those cases where Armenian hamster monoclonals need to be detected in the presence of mouse or rat immunoglobulins.
For technical information about probes, click here.
All prices are per standard unit size, and the nine-digit number is the catalog code number.
For a complete description of the product, please use the standard format to avoid mistakes when placing an order.