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 |
|---|---|---|---|
| 53049SF | 100 µg |
|
| REACTIVITY | H |
| SENSITIVITY | Endogenous |
| MW (kDa) | 30 |
| Source/Isotype | Rabbit IgG |
Product Information
Human
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro83 of human ASCL1 protein.
Achaete-scute homolog 1 (ASCL1), also known as MASH-1, is a basic helix-loop-helix (BHLH) transcription factor which plays an essential role in the differentiation of neuroendocrine cells and neural tissues (1-3). ASCL1 directly binds the E-box motif (5'-CANNTG-3') on promoters, with dimerization with other BHLH proteins required for efficient DNA binding (4). Acting as a pioneer transcription factor, ASCL1 also accesses closed chromatin, allowing other factors to promote transcription of neuronal genes and activate neural pathways (5). Research studies have shown that knockdown of the ASCL1 gene leads to inhibition of growth and induction of apoptosis in SCLC cells in vitro (6). Additionally, ASCL1 is overexpressed in both classic SCLC as well as NSCLC with neuroendocrine features, suggesting its role in the pathogenesis of those malignancies (7). ASCL1 plays a crucial role in promoting SCLC carcinogenesis by upregulating the expression of DLL3, a nonfunctioning Notch ligand, leading to inhibition of the Notch signaling pathway (8).
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