Na,K-ATPase α1 (D4Y7E) 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 critical factor.
Animal-free Manufacturing
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.
Na,K-ATPase α1 (D4Y7E) Rabbit mAb (BSA and Azide Free) #99935
Western blot analysis of extracts from SK-MEL-28, SNB19 and A-204 cells using Na,K-ATPase α1 (D4Y7E) Rabbit mAb (upper) or α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower). Data were generated using the standard formulation of this product.
Immunohistochemical analysis of paraffin-embedded SK-MEL-28 (left) and A-204 (right) cell pellets using Na,K-ATPase α1 (D4Y7E) Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemical analysis of paraffin-embedded human appendix using Na,K-ATPase α1 (D4Y7E) Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right). Data were generated using the standard formulation of this product.
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using Na,K-ATPase α1 (D4Y7E) Rabbit mAb. Data were generated using the standard formulation of this product.
Confocal immunofluorescent analysis of SK-MEL-28 (left), SNB19 (middle) or A-204 (right) cells using Na,K-ATPase α1 (D4Y7E) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). Data were generated using the standard formulation of this product.
Na,K-ATPase α1 (D4Y7E) Rabbit mAb (BSA and Azide Free) 99935
99935SF 100 µg
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REACTIVITY H
SENSITIVITY Endogenous
MW (kDa) 100
Source/Isotype Rabbit IgG
Application Key:
WB-Western Blot
IP-Immunoprecipitation
IHC-Immunohistochemistry
ChIP-Chromatin Immunoprecipitation
C&R-CUT&RUN
C&T-CUT&Tag
DB-Dot Blot
eCLIP-eCLIP
IF-Immunofluorescence
F-Flow Cytometry
Species Cross-Reactivity Key:
H-Human
M-Mouse
R-Rat
Hm-Hamster
Mk-Monkey
Vir-Virus
Mi-Mink
C-Chicken
Dm-D. melanogaster
X-Xenopus
Z-Zebrafish
B-Bovine
Dg-Dog
Pg-Pig
Sc-S. cerevisiae
Ce-C. elegans
Hr-Horse
GP-Guinea Pig
Rab-Rabbit
All-All Species Expected
Product Usage Information
This product is the carrier free version of product #23565. 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 #23565
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.
Na,K-ATPase α1 (D4Y7E) Rabbit mAb (BSA and Azide Free) recognizes endogenous levels of total Na,K-ATPase α1 protein.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro12 of human Na,K-ATPase α1 protein.
The Na,K-ATPase is an integral membrane heterodimer belonging to the P-type ATPase family. This ion channel uses the energy derived from ATP hydrolysis to maintain membrane potential by driving sodium export and potassium import across the plasma membrane against their electrochemical gradients. It is composed of a catalytic α subunit and a β subunit (reviewed in 1). Several phosphorylation sites have been identified for the α1 subunit. Tyr10 is phosphorylated by an as yet undetermined kinase (2), Ser16 and Ser23 are phosphorylated by PKC, and Ser943 is phosphorylated by PKA (3-5). All of these sites have been implicated in the regulation of enzyme activity in response to hormones and neurotransmitters, altering trafficking and kinetic properties of Na,K-ATPase. Altered phosphorylation in response to angiotensin II stimulates activity in the rat proximal tubule (6). Na,K-ATPase is also involved in other signal transduction pathways. Insulin regulates its localization in differentiated primary human skeletal muscle cells, and this regulation is dependent on ERK1/2 phosphorylation of the α subunit (7). Na,K-ATPase and Src form a signaling receptor complex that affects regulation of Src kinase activity and, subsequently, its downstream effectors (8,9).
Therien, A.G. and Blostein, R. (2000) Am J Physiol Cell Physiol 279, C541-66.
Féraille, E. et al. (1999) Mol Biol Cell 10, 2847-59.
Fisone, G. et al. (1994) J Biol Chem 269, 9368-73.
Feschenko, M.S. and Sweadner, K.J. (1995) J Biol Chem 270, 14072-7.
Beguin, P. et al. (1994) J Biol Chem 269, 24437-45.
Yingst, D.R. et al. (2004) Am J Physiol Renal Physiol 287, F713-21.
Al-Khalili, L. et al. (2004) J Biol Chem 279, 25211-8.
Tian, J. et al. (2006) Mol Biol Cell 17, 317-26.
Liang, M. et al. (2006) J Biol Chem 281, 19709-19.
Select Your Pathway/Protein Protein: P05023
Entrez-Gene Id: 476
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