Phospho-AMPKα (Thr172) (40H9) Rabbit mAb (BSA and Azide Free)
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.
Phospho-AMPKα (Thr172) (40H9) Rabbit mAb (BSA and Azide Free) #74281
This product is the carrier free version of product #2535. 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 #2535
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
Phospho-AMPKα (Thr172) (40H9) Rabbit mAb (BSA and Azide Free) detects endogenous AMPKα only when phosphorylated at threonine 172. The antibody detects both α1 and α2 isoforms of the catalytic subunit, but does not detect the regulatory β or γ subunits.
Human, Mouse, Rat, Hamster, Monkey, D. melanogaster, S. cerevisiae
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.
Chicken, Zebrafish, Bovine, Pig
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Thr172 of human AMPKα protein.
AMP-activated protein kinase (AMPK) is highly conserved from yeast to plants and animals and plays a key role in the regulation of energy homeostasis (1). AMPK is a heterotrimeric complex composed of a catalytic α subunit and regulatory β and γ subunits, each of which is encoded by two or three distinct genes (α1, 2; β1, 2; γ1, 2, 3) (2). The kinase is activated by an elevated AMP/ATP ratio due to cellular and environmental stress, such as heat shock, hypoxia, and ischemia (1). The tumor suppressor LKB1, in association with accessory proteins STRAD and MO25, phosphorylates AMPKα at Thr172 in the activation loop, and this phosphorylation is required for AMPK activation (3-5). AMPKα is also phosphorylated at Thr258 and Ser485 (for α1; Ser491 for α2). The upstream kinase and the biological significance of these phosphorylation events have yet to be elucidated (6). The β1 subunit is post-translationally modified by myristoylation and multi-site phosphorylation including Ser24/25, Ser96, Ser101, Ser108, and Ser182 (6,7). Phosphorylation at Ser108 of the β1 subunit seems to be required for AMPK activation, while phosphorylation at Ser24/25 and Ser182 affects AMPK localization (7). Several mutations in AMPKγ subunits have been identified, most of which are located in the putative AMP/ATP binding sites (CBS or Bateman domains). Mutations at these sites lead to reduction of AMPK activity and cause glycogen accumulation in heart or skeletal muscle (1,2). Accumulating evidence indicates that AMPK not only regulates the metabolism of fatty acids and glycogen, but also modulates protein synthesis and cell growth through EF2 and TSC2/mTOR pathways, as well as blood flow via eNOS/nNOS (1).