Dihydrofolate reductase Recombinant Rabbit Monoclonal Antibody [JE31-05]
Catalog# HA721394
Dihydrofolate reductase Recombinant Rabbit Monoclonal Antibody [JE31-05]
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WB
-
IHC-P
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Human
-
Mouse
-
Rat
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unconjugated
Overview
Product Name
Dihydrofolate reductase Recombinant Rabbit Monoclonal Antibody [JE31-05]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within Human Dihydrofolate reductase aa 88-187 / 187.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IHC-P
Target Molecular Weight
Predicted band size: 21.5 kDa
Positive Control
HCT116 cell lysate, HeLa cell lysate, U-2 OS cell lysate, K-562 cell lysate, MCF7 cell lysate, HEK-293 cell lysate, Jurkat cell lysate, HepG2 cell lysate, LO2 cell lysate, mouse liver tissue lysate, rat liver tissue lysate, human colon carcinoma tissue, human liver tissue.
Conjugation
unconjugated
Clone Number
JE31-05
RRID
Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
-
WB
-
1:1,000
-
IHC-P
-
1:200
Target
Function
Dihydrofolate reductase, or DHFR, is an enzyme that reduces dihydrofolic acid to tetrahydrofolic acid, using NADPH as an electron donor, which can be converted to the kinds of tetrahydrofolate cofactors used in 1-carbon transfer chemistry. In humans, the DHFR enzyme is encoded by the DHFR gene. It is found in the q14.1 region of chromosome 5. There are two structural classes of DHFR, evolutionarily unrelated to each other. The former is usually just called DHFR and is found in bacterial chromosomes and animals. In bacteria, however, antibiotic pressure has caused this class to evolve different patterns of binding diaminoheterocyclic molecules, leading to many "types" named under this class, while mammalian ones remain highly similar. The latter (type II), represented by the plastid-encoded R67, is a tiny enzyme that works by forming a homotetramer. Dihydrofolate reductase converts dihydrofolate into tetrahydrofolate, a proton shuttle required for the de novo synthesis of purines, thymidylic acid, and certain amino acids. While the functional dihydrofolate reductase gene has been mapped to chromosome 5, multiple intronless processed pseudogenes or dihydrofolate reductase-like genes have been identified on separate chromosomes. Found in all organisms, DHFR has a critical role in regulating the amount of tetrahydrofolate in the cell. Tetrahydrofolate and its derivatives are essential for purine and thymidylate synthesis, which are important for cell proliferation and cell growth. DHFR plays a central role in the synthesis of nucleic acid precursors, and it has been shown that mutant cells that completely lack DHFR require glycine, a purine, and thymidine to grow. DHFR has also been demonstrated as an enzyme involved in the salvage of tetrahydrobiopterin from dihydrobiopterin.
Background References
1. El-Subbagh HI et al. 2-Substituted-mercapto-quinazolin-4(3H)-ones as DHFR Inhibitors. Mini Rev Med Chem. 2021
2. Wróbel A et al. Recent Design and Structure-Activity Relationship Studies on the Modifications of DHFR Inhibitors as Anticancer Agents. Curr Med Chem. 2021
Subcellular Location
Mitochondrion, Cytoplasm.
Synonyms
DHFR antibody
DHFRP1 antibody
Dihydrofolate reductase antibody
DYR antibody
DYR_HUMAN antibody
EC 1.5.1.3 antibody
Images
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Western blot analysis of Dihydrofolate reductase on different lysates with Rabbit anti-Dihydrofolate reductase antibody (HA721394) at 1/1,000 dilution.
Lane 1: HCT116 cell lysate (15 µg/Lane)
Lane 2: HeLa cell lysate (15 µg/Lane)
Lane 3: U-2 OS cell lysate (15 µg/Lane)
Lane 4: K-562 cell lysate (15 µg/Lane)
Lane 5: MCF7 cell lysate (15 µg/Lane)
Lane 6: HEK-293 cell lysate (15 µg/Lane)
Lane 7: Jurkat cell lysate (15 µg/Lane)
Lane 8: HepG2 cell lysate (15 µg/Lane)
Lane 9: LO2 cell lysate (15 µg/Lane)
Lane 10: Mouse liver tissue lysate (30 µg/Lane)
Lane 11: Rat liver tissue lysate (30 µg/Lane)
Predicted band size: 21.5 kDa
Observed band size: 21.5 kDa
Exposure time: 1 minute;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721394) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature. -
Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-Dihydrofolate reductase antibody (HA721394) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721394) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-Dihydrofolate reductase antibody (HA721394) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721394) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"