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Anti-Dihydrofolate reductase (DHFR) Rabbit Antibody [M17P5]

Catalog No.: F3966

Application: Reactivity: Human, Mouse, Rat

Usage Information

Dilution
WB1:2000 IHC1:100

Application
WB, IP, IHC

Reactivity
Human, Mouse, Rat

Source
Rabbit

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW Observed MW
21 kDa 22 kDa
^*Why do the predicted and actual molecular weights differ? The following reasons may explain differences between the predicted and actual protein molecular weight.

Positive Control	Human breast cancer; Human liver cancer; Mouse liver; Mouse kidney; Rat heart; HeLa; Jurkat; K-562; Raji; HEK-293T
Negative Control

Datasheet & SDS

Biological Description

Specificity
Dihydrofolate reductase (DHFR) Rabbit mAb detects endogenous levels of total Dihydrofolate reductase (DHFR) protein.

Clone
M17P5

Synonym(s)
Dihydrofolate reductase, DHFR

Background
Dihydrofolate reductase (DHFR) is a ubiquitously expressed NADPH-dependent oxidoreductase, a member of the reductase enzyme family that catalyzes the reduction of dihydrofolate (DHF) to tetrahydrofolate (THF), an essential cofactor for one-carbon transfer reactions in purine, thymidylate, and methionine synthesis, thereby supporting DNA synthesis, repair, and methylation. Structurally, DHFR adopts a conserved α/β-fold consisting of an eight-stranded β-sheet flanked by α-helices, with a dynamic Met20 loop that regulates substrate binding and product release at the active site, where NADPH donates a hydride to DHF. It is expressed in all proliferating cells, with transcription regulated by factors such as Sp1 and E2F, peaking at the G1/S phase of the cell cycle. Functionally, DHFR maintains folate metabolism, supports homocysteine remethylation through 5-methyl-THF production, and is the primary target of antifolate drugs such as methotrexate in cancer and inflammatory disease therapy. Genetic polymorphisms, such as the 19-bp intronic deletion, can alter its expression and activity, influencing susceptibility to neural tube defects, cancer, and antifolate drug response.

Background

Dihydrofolate reductase (DHFR) is a ubiquitously expressed NADPH-dependent oxidoreductase, a member of the reductase enzyme family that catalyzes the reduction of dihydrofolate (DHF) to tetrahydrofolate (THF), an essential cofactor for one-carbon transfer reactions in purine, thymidylate, and methionine synthesis, thereby supporting DNA synthesis, repair, and methylation. Structurally, DHFR adopts a conserved α/β-fold consisting of an eight-stranded β-sheet flanked by α-helices, with a dynamic Met20 loop that regulates substrate binding and product release at the active site, where NADPH donates a hydride to DHF. It is expressed in all proliferating cells, with transcription regulated by factors such as Sp1 and E2F, peaking at the G1/S phase of the cell cycle. Functionally, DHFR maintains folate metabolism, supports homocysteine remethylation through 5-methyl-THF production, and is the primary target of antifolate drugs such as methotrexate in cancer and inflammatory disease therapy. Genetic polymorphisms, such as the 19-bp intronic deletion, can alter its expression and activity, influencing susceptibility to neural tube defects, cancer, and antifolate drug response.

References
https://pubmed.ncbi.nlm.nih.gov/21629435/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

Tel: +1-832-582-8158 Ext:3
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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%