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Human/Mouse/Rat Dihydrofolate Reductase/DHFR Antibody

Catalog #: MAB7934
5 Citations Datasheet / COA / SDS
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MAB7934
MAB7934-SP
Detection of Human, Mouse, and Rat Dihydrofolate Reductase/DHFR by Western Blot.
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Citations (5)
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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human/Mouse/Rat Dihydrofolate Reductase/DHFR Antibody Summary

Species Reactivity
Human, Mouse, Rat
Specificity
Detects human Dihydrofolate Reductase/DHFR in ELISA. Detects human, mouse and rat Dihydrofolate Reductase in Western Blots.
Source
Monoclonal Mouse IgG2B Clone # 872442
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
E. coli-derived recombinant human Dihydrofolate Reductase/DHFR
Met1-Asp187
Accession # P00374
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.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
See below
Simple Western
10 µg/mL
See below
Immunohistochemistry
8-25 µg/mL
See below
Immunocytochemistry
8-25 µg/mL
See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Human, Mouse, and Rat Dihydrofolate Reductase/DHFR antibody by Western Blot. View Larger

Detection of Human, Mouse, and Rat Dihydrofolate Reductase/DHFR by Western Blot. Western blot shows lysates of 293T human embryonic kidney cell line, Raji human Burkitt's lymphoma cell line, C2C12 mouse myoblast cell line, Rat-2 rat embryonic fibroblast cell line. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Dihydrofolate Reductase/DHFR Monoclonal Antibody (Catalog # MAB7934) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). 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.

Immunocytochemistry Dihydrofolate Reductase/DHFR antibody in MCF-7 Human Cell Line by Immunocytochemistry (ICC). View Larger

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 Mouse Anti-Human Dihydrofolate Reductase/DHFR Monoclonal Antibody (Catalog # MAB7934) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to the cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Immunocytochemistry Dihydrofolate Reductase/DHFR antibody in C2C12 Mouse Cell Line by Immunocytochemistry (ICC). View Larger

Dihydrofolate Reductase/DHFR in C2C12 Mouse Cell Line. Dihydrofolate Reductase/DHFR was detected in immersion fixed C2C12 mouse myoblast cell line using Mouse Anti-Human Dihydrofolate Reductase/DHFR Monoclonal Antibody (Catalog # MAB7934) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Immunohistochemistry Dihydrofolate Reductase/DHFR antibody in Human Colon by Immunohistochemistry (IHC-P). View Larger

Dihydrofolate Reductase/DHFR in Human Colon. Dihydrofolate Reductase/DHFR was detected in formalin fixed paraffin-embedded sections of human colon tissue using Mouse Anti-Human Dihydrofolate Reductase/DHFR Monoclonal Antibody (Catalog # MAB7934) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to the cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Simple Western Detection of Human Dihydrofolate Reductase/DHFR antibody by Simple Western<sup>TM</sup>. View Larger

Detection of Human Dihydrofolate Reductase/DHFR by Simple WesternTM. Simple Western lane view shows lysates of 293T human embryonic kidney cell line, loaded at 0.5 mg/mL. A specific band was detected for Dihydrofolate Reductase/DHFR at approximately 28 kDa (as indicated) using 10 µg/mL of Mouse Anti-Human/Mouse/Rat Dihydrofolate Reductase/DHFR Monoclonal Antibody (Catalog # MAB7934). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system. Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
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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.

Background: Dihydrofolate Reductase/DHFR

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.

Entrez Gene IDs
1719 (Human); 13361 (Mouse); 24312 (Rat)
Alternate Names
DHFR; DHFRP1; Dihydrofolate Reductase; DYR; EC 1.5.1.3

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Citations for Human/Mouse/Rat Dihydrofolate Reductase/DHFR Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

5 Citations: Showing 1 - 5
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  1. The AAA ATPase Afg1 preserves mitochondrial fidelity and cellular health by maintaining mitochondrial matrix proteostasis
    Authors: Edward M. Germany, Nataliya Zahayko, Mason L. Huebsch, Jennifer L. Fox, Veena Prahlad, Oleh Khalimonchuk
    Journal of Cell Science
  2. Uncoupled nitric oxide synthase activity promotes colorectal cancer progression
    Authors: Asim Alam, Steven C. Smith, Sundaresan Gobalakrishnan, Mina McGinn, Vasily A. Yakovlev, Christopher S. Rabender
    Frontiers in Oncology
  3. Characterization of newly established Pralatrexate-resistant cell lines and the mechanisms of resistance
    Authors: K Oiwa, N Hosono, R Nishi, L Scotto, OA O'Connor, T Yamauchi
    BMC Cancer, 2021-07-31;21(1):879.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  4. Cross Talk between One-Carbon Metabolism, Eph Signaling, and Histone Methylation Promotes Neural Stem Cell Differentiation
    Authors: MA Fawal, T Jungas, A Kischel, C Audouard, JS Iacovoni, A Davy
    Cell Rep, 2018-06-05;23(10):2864-2873.e7.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot
  5. Reprogramming of nucleotide metabolism by interferon confers dependence on the replication stress response pathway in pancreatic cancer cells
    Authors: Evan R. Abt, Thuc M. Le, Amanda M. Dann, Joseph R. Capri, Soumya Poddar, Vincent Lok et al.
    Cell Reports

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