Recombinant antibodies offer several key advantages compared to traditional antibodies. These include superior lot-to-lot consistency, continuous supply, and animal-free manufacturing. As such, recombinant antibodies are seeing increased use for scientific research, especially as a means of addressing the ongoing reproducibility crisis.
Traditional polyclonal and monoclonal antibodies are the product of normal B cell development and genetic recombination. They are generated by immunizing an animal with an antigen to elicit an immune response. While polyclonal antibodies are secreted by many different B cell clones and recognize multiple antigenic epitopes, monoclonals originate from a single B cell clone and are specific for just one epitope.
Recombinant antibodies are monoclonal, but their production involves in vitro genetic manipulation. After cloning the antibody genes into an expression vector, this is then transfected into an appropriate host cell line for antibody expression. Mammalian cell lines are most commonly used for recombinant antibody production, although cell lines of bacterial, yeast, or insect origin are also suitable.
Because recombinant antibody production involves sequencing the antibody light and heavy chains, it is a highly controlled and reliable process. In contrast, hybridoma-based systems for producing monoclonal antibodies are subject to genetic drift and instability, increasing the potential for lot-to-lot variability or loss of antibody expression. Recombinant antibodies are highly consistent from lot to lot, thereby ensuring reproducible experimental results.
In vitro methods for producing antibodies are amenable to large-scale production, meaning antibody availability is unlikely to become a limiting factor. Moreover, since the recombinant antibody sequence is known, continuity of supply is assured; in situations where an antibody will be used to support large, long-term studies, this can be an especially critical factor.
Unlike traditional methods for antibody production, recombinant approaches avoid the need to use animals. Where polyclonal antibodies are purified directly from the serum of the immunized host, and monoclonals are purified from either hybridoma-derived tissue culture supernatant or ascites, recombinant antibodies are instead purified from the tissue culture supernatants of transfected host cell lines. Regardless of whether an antibody is polyclonal, monoclonal or recombinant, it must always be properly validated in the intended application prior to experimental use. At CST, we adhere to the Hallmarks of Antibody Validation™, six complementary strategies for determining the specificity, sensitivity, and functionality of an antibody in any given assay. By carefully tailoring these strategies to each antibody product, we guarantee that CST antibodies will work as expected, to help you achieve results you can trust.
| Cat. # | Size | Qty. | Price |
|---|---|---|---|
| 93671S | 100 µl (50 tests) |
|
| Concentration | 100 ug/ml |
| SENSITIVITY | Endogenous |
| Source/Isotype | Rabbit IgG |
Product Information
This Cell Signaling Technology® F(ab’)2 fragment lacks an Fc domain, and will not exhibit reactivity with human Fc receptors. This eliminates the need for Fc blocking and reduces false positives in assays containing human monocytes or other cells with high expression of Fc receptors.
The F(ab')2 fragment is generated by enzymatic digestion of the full-length IgG clone coupled with chromatography to remove both the Fc fragment and the intact IgG. The F(ab’)2 fragment is conjugated to Alexa Fluor® 647 fluorescent dye under optimal conditions.
Rabbit (DA1E) IgG XP® Isotype Control F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) is not directed against any known antigen. It functions as an isotype control for rabbit IgG antibodies. This F(ab')2 fragment contains no Fc domain and will not exhibit cross-reactivity with Fc receptors.
Isotype control antibodies are used to estimate the nonspecific binding of target primary antibodies due to Fc receptor binding or other protein-protein interactions. An isotype control antibody should have the same immunoglobulin type and be used at the same concentration as the test antibody.
Human Fc receptors exhibit strong cross-reactivity with rabbit IgG, and are expressed at high levels on cells of myeloid lineage (e.g., monocytes and myeloid-derived cell lines). Fluorescent F(ab')2 fragment conjugates are ideal for use in flow cytometry assays on cells containing Fc receptors, and eliminate the need for an Fc blocking step prior to antibody incubation.
Cell Signaling Technology employs strict quality control procedures to ensure that each conjugate provides optimal specificity and fluorescence.
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