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.
β-Actin (D6A8) Rabbit mAb (BSA and Azide Free) #19069
This product is the carrier free version of product #8457. 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 #8457
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
β-Actin (D6A8) Rabbit mAb (BSA and Azide Free) recognizes endogenous levels of total β-actin protein. Due to the high sequence identity between the cytoplasmic actin isoforms, β-actin and cytoplasmic γ-actin, this antibody may cross-react with cytoplasmic γ-actin. It does not cross-react with α-skeletal, α-cardiac, α-vascular smooth, or γ-enteric smooth muscle isoforms.
Human, Mouse, Rat, Monkey, D. melanogaster, Zebrafish
Species predicted to react based on 100% sequence homology:
The antigen sequence used to produce this antibody shares
100% sequence homology with the species listed here, but
reactivity has not been tested or confirmed to work by CST.
Use of this product with these species is not covered under
Antibody Performance Guarantee.
Hamster, Chicken, Bovine, Pig
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human β-actin protein.
Actin, a ubiquitous eukaryotic protein, is the major component of the cytoskeleton. At least six isoforms are known in mammals. Nonmuscle β- and γ-actin, also known as cytoplasmic actin, are ubiquitously expressed, controlling cell structure and motility (1). While all actin isoforms are highly homologous, cytoplasmic β- and γ-actin protein sequences differ by only four biochemically similar amino acids (2). For this reason, antibodies raised to β-actin may cross-react with γ-actin, and vice versa. α-cardiac and α-skeletal actin are expressed in striated cardiac and skeletal muscles, respectively; two smooth muscle actins, α- and γ-actin, are found primarily in vascular smooth muscle and enteric smooth muscle, respectively. These actin isoforms regulate the contractile potential of muscle cells (1). Actin exists mainly as a fibrous polymer, F-actin. In response to cytoskeletal reorganizing signals during processes such as cytokinesis, endocytosis, or stress, cofilin promotes fragmentation and depolymerization of F-actin, resulting in an increase in the monomeric globular form, G-actin (3). The ARP2/3 complex stabilizes F-actin fragments and promotes formation of new actin filaments (3). Research studies have shown that actin is hyperphosphorylated in primary breast tumors (4). Cleavage of actin under apoptotic conditions has been observed in vitro and in cardiac and skeletal muscle, as shown in research studies (5-7). Actin cleavage by caspase-3 may accelerate ubiquitin/proteasome-dependent muscle proteolysis (7).