DHFR Recombinant Monoclonal Antibody

Code CSB-RA264582A0HU
Size US$210
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Image
  • Western Blot
    Positive WB detected in: Hela whole cell lysate, Jurkat whole cell lysate, K562 whole cell lysate, Raji whole cell lysate, 293T whole cell lysate, Rat heart tissue, Mouse kidney tissue, Mouse liver tissue
    All lanes: DHFR antibody at 1:2000
    Secondary
    Goat polyclonal to rabbit IgG at 1/50000 dilution
    Predicted band size: 22, 16 kDa
    Observed band size: 22 kDa
  • IHC image of CSB-RA264582A0HU diluted at 1:100 and staining in paraffin-embedded human breast cancer performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4℃ overnight. The primary is detected by a Goat anti-rabbit IgG polymer labeled by HRP and visualized using 0.05% DAB.
  • IHC image of CSB-RA264582A0HU diluted at 1:100 and staining in paraffin-embedded human liver cancer performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4℃ overnight. The primary is detected by a Goat anti-rabbit IgG polymer labeled by HRP and visualized using 0.05% DAB.
  • Immunoprecipitating DHFR in Hela whole cell lysate
    Lane 1: Rabbit control IgG instead of CSB-RA264582A0HU in Hela whole cell lysate. For western blotting,a HRP-conjugated Protein G antibody was used as the secondary antibody (1/2000)
    Lane 2: CSB-RA264582A0HU(2µg)+ Hela whole cell lysate(500µg)
    Lane 3: Hela whole cell lysate (10µg)
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Product Details

Uniprot No.
Target Names
DHFR
Alternative Names
Dihydrofolate reductase (EC 1.5.1.3), DHFR
Species Reactivity
Human, Mouse, Rat
Immunogen
A synthesized peptide derived from human DHFR
Immunogen Species
Homo sapiens (Human)
Conjugate
Non-conjugated
Clonality
Monoclonal
Isotype
Rabbit IgG
Clone No.
9B2
Purification Method
Affinity-chromatography
Concentration
It differs from different batches. Please contact us to confirm it.
Buffer
Rabbit IgG in phosphate buffered saline, pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.
Form
Liquid
Tested Applications
ELISA, WB, IHC, IP
Recommended Dilution
Application Recommended Dilution
WB 1:500-1:5000
IHC 1:50-1:200
IP 1:200-1:1000
Troubleshooting and FAQs
Storage
Upon receipt, store at -20°C or -80°C. Avoid repeated freeze.
Lead Time
Basically, we can dispatch the products out in 1-3 working days after receiving your orders. Delivery time maybe differs from different purchasing way or location, please kindly consult your local distributors for specific delivery time.
Description

The DNA sequence corresponding to the DHFR monoclonal antibody produced from the animals through the human DHFR synthesized peptide immunization was cloned into the expression vector, which was further transfected into a cell line for in vitro expression. The product is the recombinant DHFR monoclonal antibody. It specifically targets the DHFR from human, mouse, and rat species. It belongs to the rabbit IgG. The affinity-chromatography purification method was used to purify this DHFR antibody. This GHFR recombinant antibody has been tested by ELISA, WB, IHC, and IP to assure specificity and reactivity.

DHFR, an oxidoreductase, catalyzes the reduction of FH2acid to H4FA, thus regulating the regeneration of H4FA. DHFR has been demonstrated to react with the Hedgehog (Hh) signaling pathway negative regulator SuFu. Abnormal activation of the Hh signaling pathway is closely related to the occurrence and development of tumors. This means that DHFR knockdown and overexpression affect the proliferation and apoptosis of tumor cells.

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Target Background

Function
Key enzyme in folate metabolism. Contributes to the de novo mitochondrial thymidylate biosynthesis pathway. Catalyzes an essential reaction for de novo glycine and purine synthesis, and for DNA precursor synthesis. Binds its own mRNA and that of DHFR2.
Gene References into Functions
  1. In ALL and NHL patients treated with methotrexate, treatment toxicities and outcome were evaluated. Multivariate analysis showed that DHFR-1610G/T (OR=0.107, p=0.018) and MTHFR677T alleles (OR=0.12, p=0.026) had a strong protective effect against hematologic toxicity, while DHFR-1610CC genotype increased this toxicity (OR=9, p=0.045). PMID: 28887233
  2. Dihydrofolate reductase and thymidylate synthase form a complex in vitro and co-localize in normal and cancer cells. PMID: 27187663
  3. Single nucleotide polymorphism in DHFR gene is associated with Systemic lupus erythematosus. PMID: 28943344
  4. study concludes polymorphism 63/91 in DHFR gene promoter can modulate the onset of methotrexate-related adverse effects in rheumatoid arthritis patients PMID: 27636122
  5. our findings suggest that the identification of DHFR polymorphisms in the promoter region of the gene may be helpful in tailoring MTX doses for ALL pediatric patients on maintenance therapy. PMID: 28719513
  6. The abundance of dihydrofolate reductase was statistically significantly increased in rheumatoid arthritis (RA)-patient biopsies compared with controls and correlated with the administered dosage of methotrexate (MTX), the most frequently prescribed immunosuppressive drug for RA. PMID: 27627584
  7. The present study demonstrated that ADAR1 positively regulates the expression of DHFR by editing the miR-25-3p and miR-125a-3p binding sites in the 3'-UTR of DHFR, enhancing cellular proliferation and resistance to methotrexate in MCF-7 cells. PMID: 28188287
  8. In conclusion, the finding suggests that folate nutrition and 19bp del-DHFR [Dihydrofolate reductase] variation may interact to modify adenomatous polyp [colorectal cancer] risk. PMID: 26875486
  9. the highest expression of GGH and EGFR was noted in the left-sided colon; the highest expression of DHFR, FPGS, TOP1 and ERCC1 was noted in the rectosigmoid, whereas TYMP expression was approximately equivalent in the right-sided colon and rectum PMID: 26676887
  10. Overexpression of miR-192 inhibited cellular proliferation by binding DHFR. miR-192 decreased cellular anchoring via the repression of ITGAV, ITGB1, ITGB3, and CD47 PMID: 26506238
  11. Data suggest that DHFR exhibits intrinsic activity kinetics that are temperature-independent; additional mass (i.e., incorporation of H, C, and N isotopes) has no effect on intrinsic activity kinetics or protein conformation/stability of DHFR. PMID: 26813442
  12. patients homozygous for the G allele of rs1053129 in the DHFR gene were more likely to have a metastasis (45%, P= 0.005), and the methylenetetetrahydrofolate reductase (MTHFR) 677C allele was associated with higher degree of liver toxicity PMID: 25778468
  13. the association between cognitive outcomes with the 19-bp deletion DHFR polymorphism, folate status, and their interaction with high or normal plasma folate PMID: 26354538
  14. S-nitrosylation of DHFR at cysteine 7 by eNOS-derived NO is crucial for DHFR stability. PMID: 26381869
  15. genetic association studies in cohort of healthy young adults in Ireland: Data suggest that a 19 bp deletion/insertion polymorphism within intron 1 of DHFR (rs70991108) is not associated with folate nutritional status in the population studied. PMID: 26269242
  16. Dihydrofolate Reductase and Thymidylate Synthase Transgenes Resistant to Methotrexate Interact to Permit Novel Transgene Regulation. PMID: 26242737
  17. results for the first time suggested the DHFR 19-bp D/D genotype may confer a reduced risk of NS-CL/P and might act as a protective factor against NS-CL/P in the Iranian subjects. PMID: 26221921
  18. Triple mutant haplotypes AIRNI (N51I+C59R+S108N) of the dhfr gene associated with pyrimethamine resistance were prevalent in Chirang district of Assam. PMID: 25511211
  19. The human dihydrofolate reductase is relatively less stable than its E.coli counterpart. PMID: 26349210
  20. subpicosecond protein motion is dynamically coupled to hydride transfer catalyzed by hsDHFR but not ecDHFR PMID: 25369552
  21. MTHFR, DHFR and ATIC genetic variants can be considered as pharmacogenetic markers of outcome in RA patients under MTX monotherapy. PMID: 25084201
  22. Genome-wide association studies identify 10 novel genetic loci as risk factors for methotrexate response in rheumatoid arthritis patients additionally to polymorphism in DHFR, FPGS and TYMS. PMID: 24583629
  23. Genetic association between DHFR single nucleotide polymorphisms and nonsyndromic cleft lip with or without cleft palate. PMID: 24361572
  24. The hDHFR requires minimal backbone conformational rearrangement as it proceeds through its enzymatic cycle, but that ligand flux is brokered by more subtle conformational changes that depend on the side chain motions of critical residues. PMID: 24498949
  25. berberine suppresses the growth of cDDP-resistant cells more than the sensitive counterparts, by interfering with the expression of folate cycle enzymes, dihydrofolate reductase (DHFR) and thymidylate synthase (TS). PMID: 23903781
  26. Data suggest that methylenetetrahydrofolate reductase (NAD(P)H), dihydrofolate reductase, thymidylate synthetase and SLC19A1 genes present increased expression after the highest dose of methotrexate in laryngeal cancer cell line. PMID: 23838799
  27. Despite structural similarity, Escherichia coli and human DHFRs use different dynamic mechanisms to perform the same function, and human DHFR cannot complement DHFR-deficient E. coli cells. PMID: 24077226
  28. Interactions between the ligands and Asn 64, Phe 31, and Phe 34 are important for increased affinity for DHFR. PMID: 24053334
  29. We conclude that low activity of endothelial DHFR is an important factor limiting the benefits of BH4 therapies, which may be further aggravated by folate supplements. PMID: 23707606
  30. There is an association between DHFR DD/SHMT TT and DHFR II/SHMT TT combined genotypes and folate and MMA concentrations in individuals with Down syndrome. PMID: 23421317
  31. High DHFR immunoexpression correlated with nodal status and primary nasopharyngeal carcinoma. PMID: 23726796
  32. The data presented here provide a glimpse into the evolutionary trajectory of functional DHFR through its protein sequence space that lead to the diverged binding and catalytic properties of the E. coli and human enzymes. PMID: 23733948
  33. Inhibits cell growth through a mechanism involving downregulation of DHFR protein. PMID: 22954684
  34. Genotyping of DHFR 829C>T and GGH -401C>T was performed using a polymerase chain reaction. PMID: 22994778
  35. Led to accelerated degradation of DHFR and to inhibition of cancer cell growth. PMID: 23197646
  36. The present study was aimed to investigate the possible association between 19-base pair (bp) deletion polymorphism of the DHFR gene (rs70991108), null genotype of UGT2B17 as well as the expression level of NGX6 with the risk of breast cancer. PMID: 23053953
  37. Rheumatoid arthritis patients with DHFR-317AA genotype had less favourable response to methotrexate. PMID: 22324981
  38. human dihydrofolate reductase is bound to NADP. PMID: 22024482
  39. Constituents of the folate cycle could be involved in the etiology of idiopathic intellectual disability. PMID: 22005284
  40. This report has clearly shown that the population rate of change of resistant dhfr and dhps alleles is contingent to the sulfadoxine-pyrimethamine usage in the population in the Morogor-Mvomero district PMID: 21857842
  41. The 19-bp deletion polymorphism of DHFR gene was not a maternal risk factor for Down syndrome and was not related to variations in the concentrations of serum folate and plasma homocysteine and methylmalonic acid in the study population. PMID: 21120433
  42. The 19-base pair deletion polymorphism of DHFR was studied in Japanese. The genotype distribution was wild/wild, 11.9%; wild/deletion, 40.1%; deletion/deletion, 48.0%. Frequencies of wild type and deletion alleles were 0.32 and 0.68, respectively. PMID: 20834190
  43. Dihydrofolate reductase deficiency is associated with inborn error of metabolism. PMID: 21310276
  44. Dihydrofolate reductase deficiency due to a homozygous DHFR mutation causes megaloblastic anemia and cerebral folate deficiency leading to severe neurologic disease. PMID: 21310277
  45. the first kinetic parameters for DHFR from pjDHFR and pcDHFR with methotrexate (MTX), trimethoprim (TMP), and its potent analogue, PY957, is reported. PMID: 19196009
  46. DNA variants have a role in predisposition to disease-free survival in childhood acute lymphoblastic leukemia PMID: 19861437
  47. protein folding of dihydrofolate reductases (DHFR) from human, Escherichia coli, and Lactobacillus casei were elucidated and compared using intrinsic Trp fluorescence and fluorescence-detected ANS binding PMID: 11779239
  48. studied differences between the regulation of Plasmodium and human dihydrofolate reductases PMID: 11964483
  49. Computer modeling studies of the structural role of NADPH binding to active site mutants of human dihydrofolate reductase in complex with piritrexim PMID: 11996001
  50. Molecular mechanisms that govern translational regulation of DHFR in response to MTX. Review. PMID: 12084458

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Involvement in disease
Megaloblastic anemia due to dihydrofolate reductase deficiency (DHFRD)
Subcellular Location
Mitochondrion. Cytoplasm.
Protein Families
Dihydrofolate reductase family
Tissue Specificity
Widely expressed in fetal and adult tissues, including throughout the fetal and adult brains and whole blood. Expression is higher in the adult brain than in the fetal brain.
Database Links

HGNC: 2861

OMIM: 126060

KEGG: hsa:1719

STRING: 9606.ENSP00000396308

UniGene: Hs.592364

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