Dihydrofolate Reductase/DHFR Antibody (872442)

R&D Systems | Catalog # MAB7934

R&D Systems

Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Yeast

Applications

Validated:

Immunohistochemistry, Western Blot, Immunocytochemistry, Simple Western

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 872442
View all Dihydrofolate Reductase/DHFR Primary Antibodies »

Product Specifications

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.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Scientific Data Images for Dihydrofolate Reductase/DHFR Antibody (872442)

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

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

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

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

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 antibody by Simple WesternTM.

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.

Applications for Dihydrofolate Reductase/DHFR Antibody (872442)

Application
Recommended Usage

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed C2C12 mouse myoblast cell line and MCF‑7 human breast cancer cell line

Immunohistochemistry

8-25 µg/mL
Sample: Formalin fixed paraffin-embedded sections of human colon

Simple Western

10 µg/mL
Sample: 293T human embryonic kidney cell line

Western Blot

1 µg/mL
Sample: 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

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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

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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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.

Alternate Names

DHFR, DHFRP1, DYR

Entrez Gene IDs

1719 (Human); 13361 (Mouse); 24312 (Rat)

Gene Symbol

DHFR

UniProt

P00374 (Human)

Additional Dihydrofolate Reductase/DHFR Products

Product Documents for Dihydrofolate Reductase/DHFR Antibody (872442)

Certificate of Analysis

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Product Specific Notices for Dihydrofolate Reductase/DHFR Antibody (872442)

For research use only

Related Research Areas

Redox Enzymes

Citations for Dihydrofolate Reductase/DHFR Antibody (872442)

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Protocols

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