Anti-Human Secondary Antibodies
Choose from a handpicked selection of anti-human secondary antibodies with enzymatic conjugates like AP, Biotin or HRP or fluorescent conjugates including FITC, APC and DyLight and Atto dye families. Use the navigation on the left for anti-human IgG, IgA, IgE and IgM secondary antibodies. Browse our offer for a variety of different host, isotypes or binding specifities.
Immunoassay Production
Immunoassays are commonly utilized as diagnostic tool based on highly specific binding between antibodies and antigens. Rapid test assays like lateral flow provide qualitative results within minutes, where as an ELISA can take hours to perform but provides quantitative information.
Choosing Anti-Human Secondary Antibodies for Assay Production
While the function and purpose of immunoassays differ, high-quality reagents are essential when manufacturing serological kits for clinical use. The choice of the right Anti-Human Secondary Antibody for an assay depends on several factors. It is important to consider the species in which the primary antibody was generated and to use a secondary antibody that has minimal cross-reactivity to that species.
The type of immunoassay determines the conjugate. Secondary human antibodies linked with enzymes such as HRP and alkaline phosphatase AP are commonly utilized in applications such as ELISA or Western blotting. FITC, TRITC, APC or PE or commercial dye families as conjugate excel in flow cytometry or immunofluorescence experiments.
Anti-Human Secondary Antibodies Isotypes
In humans, there are five heavy chain isotypes α,δ,γ,ε,μ, corresponding to five antibody isotypes. Most immunoassays test for IgG isotypes, IgG is the antibody most frequently found in blood circulation, making up between 10 and 20% of total plasma proteins. IgG is also the most prevalent antibody type among the five classes, comprising around 75% of serum antibodies which are produced as part of the secondary immune response. IgM and IgA a frequently used targets for serological immune assays, they can be detected earlier in immune response which represents an advantage valuable in diagnostics. Antibodie-online offers a variety of anti-human IgG, IgA, IgE and IgM secondary antibodies. Your experimental setup requires to detect multiple immunoglobulins at once? Browse our selection at the end of this article.
Binding specificities of Anti-Human Secondary Antibodies
Secondary antibodies can target the whole immunoglobulin or only a fragment, eg the Fc, F(ab’)2 or Fab domain. Whole IgG antibodies are often sufficient, but F(ab’)2 and Fab fragments are useful to avoid binding to cells with Fc receptors.
Anti-Fc or heavy-chain (α, δ, ε, γ, and μ) IgG antibodies react with the heavy-chain only. They are specific and do not react with light-chains or heavy-chains of non IgG antibodies such as IgA, IgD, IgE and IgM.
F(ab’)2 fragment antibodies on the other hand lack most of the Fc region of the whole IgG molecule - their small size enables them to penetrate tissues easier. F(ab)2 fragment antibodies are well suited for IHC experiments, better tissue penetration improves antigen recognition and therefore immunohistochemistry signals. Since IgG F(ab’)2 fragment antibodies react with light-chains these antibodies do not exclusively bind to IgG, other immunoglobulins sharing the same light-chain as IgG are also recognized. These antibodies are mainly used in double labeling experiments or when tissues/cells containing Fc receptors are used.
In comparison to whole IgG and F(ab’)2 fragment antibodies, F(ab) fragment antibodies only have one antigen binding site. These antibodies are mainly used for blocking endogenous immunoglobulins on cells/tissues and for double labeling experiments in which researchers would like use primary antibodies from the same host species.
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