DHFR (DiHydroFolate Reductase; also Tetrahydrofolate dehydrogenase) is a 21-23 kDa member of the dihydrofolate reductase family of enzymes. It is a ubiquitously expressed monomer, and considered to be a housekeeping gene. Housekeeping genes are those that play a role in multiple pathways, although not the same pathway(s) in all cells. DHFR participates in the reduction of dihydrofolate to tetrahydrofolate, a product that is subsequently used in the synthesis of purines and thymidylic acid that are used to generate both RNA and DNA. Within the cell, DHFR is known to exist in two pools: one contains DHFR bound to its own RNA where it acts as a transcriptional repressor, while another contains DHFR bound to NADPH. Human DHFR is 187 amino acids (aa) in length and possesses one DHFR domain (aa 4-185). Its mRNA binding motif is suggested to involve Cys6, Leu22, Glu30 and Ser118. There is one potential alternative start site found 75 aa upstream of the standard start site. Full length human DHFR (aa 1-187) shares 90% aa sequence identity with mouse DHFR.
Human Dihydrofolate Reductase/DHFR Antibody
R&D Systems | Catalog # AF7934
Key Product Details
Species Reactivity
Human
Applications
Western Blot, Immunocytochemistry
Label
Unconjugated
Antibody Source
Polyclonal Sheep IgG
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Product Specifications
Immunogen
E. coli-derived recombinant human Dihydrofolate Reductase/DHFR
Met1-Asp187
Accession # P00374
Met1-Asp187
Accession # P00374
Specificity
Detects human Dihydrofolate Reductase/DHFR in direct ELISAs and Western blots.
Clonality
Polyclonal
Host
Sheep
Isotype
IgG
Scientific Data Images for Human Dihydrofolate Reductase/DHFR Antibody
Detection of Human Dihydrofolate Reductase/DHFR by Western Blot.
Western blot shows lysates of COLO 205 human colorectal adenocarcinoma cell line, HEK293T human embryonic kidney cell line, and Raji human Burkitt's lymphoma cell line. PVDF membrane was probed with 0.5 µg/mL of Sheep Anti-Human Dihydrofolate Reductase/DHFR Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7934) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Dihydrofolate Reductase/DHFR at approximately 21 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.Dihydrofolate Reductase/DHFR in MCF‑7 Human Cell Line.
Dihydrofolate Reductase/ DHFR was detected in immersion fixed MCF-7 human breast cancer cell line using Sheep Anti-Human Dihydrofolate Reductase/ DHFR Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7934) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the Northern-Lights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counter-stained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.Applications for Human Dihydrofolate Reductase/DHFR Antibody
Application
Recommended Usage
Immunocytochemistry
5-15 µg/mL
Sample: Immersion fixed MCF‑7 human breast cancer cell line
Sample: Immersion fixed MCF‑7 human breast cancer cell line
Western Blot
0.5 µg/mL
Sample: COLO 205 human colorectal adenocarcinoma cell line, HEK293T human embryonic kidney cell line, and Raji human Burkitt's lymphoma cell line
Sample: COLO 205 human colorectal adenocarcinoma cell line, HEK293T human embryonic kidney cell line, and Raji human Burkitt's lymphoma cell line
Formulation, Preparation, and Storage
Purification
Antigen Affinity-purified
Reconstitution
Sterile PBS to a final concentration of 0.2 mg/mL. For liquid material, refer to CoA for concentration.
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Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Calculators
Background: Dihydrofolate Reductase/DHFR
Alternate Names
DHFR, DHFRP1, DYR
Gene Symbol
DHFR
UniProt
Additional Dihydrofolate Reductase/DHFR Products
Product Documents for Human Dihydrofolate Reductase/DHFR Antibody
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human Dihydrofolate Reductase/DHFR Antibody
For research use only
Related Research Areas
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- ClariTSA™ Fluorophore Kits
- Detection & Visualization of Antibody Binding
- ICC Cell Smear Protocol for Suspension Cells
- ICC Immunocytochemistry Protocol Videos
- ICC for Adherent Cells
- Immunocytochemistry (ICC) Protocol
- Immunocytochemistry Troubleshooting
- Immunofluorescence of Organoids Embedded in Cultrex Basement Membrane Extract
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Fluorescent ICC Staining of Cell Smears - Graphic
- Protocol for the Fluorescent ICC Staining of Cultured Cells on Coverslips - Graphic
- Protocol for the Preparation and Fluorescent ICC Staining of Cells on Coverslips
- Protocol for the Preparation and Fluorescent ICC Staining of Non-adherent Cells
- Protocol for the Preparation and Fluorescent ICC Staining of Stem Cells on Coverslips
- Protocol for the Preparation of a Cell Smear for Non-adherent Cell ICC - Graphic
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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