Dihydrofolate Reductase/DHFR Antibody (872442) [Unconjugated]

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.

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.

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.

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 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 the12-230 kDa separation system. Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.

• Reactivity: Hu, Mu, Rt
• Applications: WB, Simple Western, IHC, ICC/IF
• Clone: 872442
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: LYOPH

Dihydrofolate Reductase/DHFR Antibody (872442) [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human Dihydrofolate Reductase/DHFR
  Met1-Asp187
  Accession # P00374
• Specificity: Detects human Dihydrofolate Reductase/DHFR in ELISA. Detects human, mouse and rat Dihydrofolate Reductase in Western Blots.
• Source: N/A
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Mouse
• Gene: DHFR
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 8-25 ug/mL
  • Immunohistochemistry 8-25 ug/mL
  • Simple Western 10 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Dihydrofolate Reductase/DHFR Antibody (872442) [Unconjugated]

• DHFR
• DHFRP1
• Dihydrofolate Reductase
• DYR
• EC 1.5.1.3

Background

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Dihydrofolate Reductase/DHFR Antibody (MAB7934)(5)

Publications using MAB7934

• Asim Alam, Steven C. Smith, Sundaresan Gobalakrishnan, Mina McGinn, Vasily A. Yakovlev, Christopher S. Rabender Uncoupled nitric oxide synthase activity promotes colorectal cancer progression Frontiers in Oncology 3/14/2023 [PMID: 36998441]
• Evan R. Abt, Thuc M. Le, Amanda M. Dann, Joseph R. Capri, Soumya Poddar, Vincent Lok et al. Reprogramming of nucleotide metabolism by interferon confers dependence on the replication stress response pathway in pancreatic cancer cells Cell Reports 1/11/2022 [PMID: 35021095]
• K Oiwa, N Hosono, R Nishi, L Scotto, OA O'Connor, T Yamauchi Characterization of newly established Pralatrexate-resistant cell lines and the mechanisms of resistance BMC Cancer, 2021-07-31;21(1):879. 2021-07-31 [PMID: 34332580] (Western Blot, Human)
• Edward M. Germany, Nataliya Zahayko, Mason L. Huebsch, Jennifer L. Fox, Veena Prahlad, Oleh Khalimonchuk The AAA ATPase Afg1 preserves mitochondrial fidelity and cellular health by maintaining mitochondrial matrix proteostasis Journal of Cell Science 5/2/2018 [PMID: 30301782]
• MA Fawal, T Jungas, A Kischel, C Audouard, JS Iacovoni, A Davy Cross Talk between One-Carbon Metabolism, Eph Signaling, and Histone Methylation Promotes Neural Stem Cell Differentiation Cell Rep, 2018-06-05;23(10):2864-2873.e7. 2018-06-05 [PMID: 29874574] (Western Blot, Mouse)

Bioinformatics

• Gene Symbol: DHFR
• Uniprot:
  • P00374(Human)