What is a Recombinant Antibody and Why is it Important?
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.
What is a Recombinant Antibody?
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.
Superior Lot-to-Lot Consistency
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
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.
53BP1 (E7N5D) XP® Rabbit mAb (BSA and Azide Free) #60721
This product is the carrier free version of product #88439. All data were generated using the same antibody clone in the standard formulation which contains BSA and glycerol.
This formulation is ideal for use with technologies requiring specialized or custom antibody labeling, including fluorophores, metals, lanthanides, and oligonucleotides. It is not recommended for ChIP, ChIP-seq, CUT&RUN or CUT&Tag assays. If you require a carrier free formulation for chromatin profiling, please contact us. Optimal dilutions/concentrations should be determined by the end user.
Supplied in 1X PBS, BSA and Azide Free.
For standard formulation of this product see product #88439
Store at -20°C. This product will freeze at -20°C so it is recommended to aliquot into single-use vials to avoid multiple freeze/thaw cycles. A slight precipitate may be present and can be dissolved by gently vortexing. This will not interfere with antibody performance.
Specificity / Sensitivity
53BP1 (E7N5D) XP® Rabbit mAb (BSA and Azide Free) recognizes endogenous levels of total 53BP1 protein.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly1190 of human 53BP1 protein.
p53-binding protein 1 (53BP1) was originally identified as a p53 binding partner that could enhance the transcriptional activity of p53 (1,2). 53BP1 consists of two BRCA1 carboxy terminal (BRCT) domains that allow for binding to p53 and a separate domain responsible for binding to phosphorylated histone H2A.X (3). 53BP1 rapidly translocates to nuclear foci following treatment of cells with ionizing radiation (IR) or radiomimetic agents that cause DNA double strand breaks (DSBs) (4,5). Because of this localization to DSBs and homology to the yeast protein Rad9, a role for 53BP1 in DSB repair has been proposed. Recruitment of 53BP1 to sites of DNA damage has been demonstrated to be independent of ATM, NBS1, and DNA-PK (4) and retention of 53BP1 at DNA breaks requires phosphorylated H2A.X (6). In cells lacking 53BP1, phosphorylation of ATM substrates is reduced, suggesting that 53BP1 is upstream of ATM (7). In response to IR, phosphorylation of 53BP1 at serines 6, 25, 29, and 784 by ATM has been demonstrated, but phosphorylation at these sites is not required for localization of 53BP1 to sites of DSBs (6). Phosphorylation of 53BP1 at Ser1618 has been reported to be enriched in human cells arrested in mitosis (8).