Human/Mouse/Rat Dihydrofolate Reductase/DHFR Antibody

  
  • 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 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.
Data Examples
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 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 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 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.
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.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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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