WB, IP, IHC-P
H M Mk
Endogenous
22
Rabbit IgG
#P00374
1719
Product Information
Product Usage Information
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
Immunoprecipitation | 1:50 |
Immunohistochemistry (Paraffin) | 1:400 - 1:1600 |
Storage
Specificity / Sensitivity
Species Reactivity:
Human, Mouse, Monkey
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly175 of human DHFR protein.
Background
Dihydrofolate reductase (DHFR) catalyzes tetrahydrofolate regeneration through the reduction of dihydrofolate using NADPH as a cofactor (1). As a key enzyme in folate metabolism, DHFR is ubiquitously expressed in the mitochondria, and is essential for the synthesis of purines, pyrimidines, and some amino acids (2). DHFR is capable of translational autoregulation by binding within the coding region of its own mRNA sequence to repress cellular DHFR protein levels (3). Mutations in the DHFR gene are known to cause inborn errors of folate metabolism resulting in megaloblastic anemia, pancytopenia, and severe cerebral folate deficiency (4). Because tetrahydrofolate is essential for DNA synthesis, cell growth, and proliferation, DHFR is a target of chemotherapeutic agents (e.g., methotrexate and pemetrexed) used in the treatment of many cancer types (5). Increased expression of DHFR has also been identified as a potential mechanism for tumor resistance to methotrexate, and therefore has been utilized as a clinical biomarker to predict patient responsiveness to folate antagonists (6,7).
- Davies, J.F. et al. (1990) Biochemistry 29, 9467-79.
- Schnell, J.R. et al. (2004) Annu Rev Biophys Biomol Struct 33, 119-40.
- Ercikan-Abali, E.A. et al. (1997) Biochemistry 36, 12317-22.
- Banka, S. et al. (2011) Am J Hum Genet 88, 216-25.
- Vander Heiden, M.G. and DeBerardinis, R.J. (2017) Cell 168, 657-69.
- Nakano, M. et al. (2017) J Biol Chem 292, 4873-84.
- Organista-Nava, J. et al. (2018) Oncol Lett 15, 8405-11.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Western Blot Buffer
IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
Applications Key
WB: Western Blotting IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin)
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
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