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Description: Cytotoxic agent
Chemical Name: N-[4-[[(2,4-Diamino-6-pteridinyl)methyl]methylamino]benzoyl]-L-glutamic acid
Purity: ≥99% (HPLC)
Citations (2)
Literature (2)

Biological Activity for Methotrexate

Methotrexate is a cytotoxic agent. Inhibits thymidylate synthetase and de novo purine synthesis. Potent folic acid antagonist; inhibits dihydrofolate reductase. Also inhibits Ras carboxyl methylation in DKOB8 cells, leading to decreased p44 and Akt activation.

Technical Data for Methotrexate

M. Wt 454.44
Formula C20H22N8O5
Storage Store at RT
Purity ≥99% (HPLC)
CAS Number 59-05-2
PubChem ID 126941
Smiles O=C(N[C@H]([C@](O)=O)CCC(O)=O)C(C=C3)=CC=C3N(CC2=NC1=C(N)N=C(N)N=C1N=C2)C

The technical data provided above is for guidance only. For batch specific data refer to the Certificate of Analysis.

Tocris products are intended for laboratory research use only, unless stated otherwise.

Solubility Data for Methotrexate

Solvent Max Conc. mg/mL Max Conc. mM
DMSO 100

Preparing Stock Solutions for Methotrexate

The following data is based on the product molecular weight 454.44. Batch specific molecular weights may vary from batch to batch due to the degree of hydration, which will affect the solvent volumes required to prepare stock solutions.

Select a batch to recalculate based on the batch molecular weight:
Concentration / Solvent Volume / Mass 1 mg 5 mg 10 mg
1 mM 2.2 mL 11 mL 22.01 mL
5 mM 0.44 mL 2.2 mL 4.4 mL
10 mM 0.22 mL 1.1 mL 2.2 mL
50 mM 0.04 mL 0.22 mL 0.44 mL

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Product Datasheets for Methotrexate

Certificate of Analysis / Product Datasheet
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References for Methotrexate

References are publications that support the biological activity of the product.

Allegra et al (1987) Evidence for direct inhibition of de novo protein synthesis in human MCF-7 breast cells as a principal mode of metabolic inhibition by methotr. J.Biol.Chem. 262 13520 PMID: 2443493

Chu et al (1990) Mechanism of thymidylate synthase inhibition by methotr. in human neoplastic cell lines and normal human myeloid progenitor cells. J.Biol.Chem. 265 8470 PMID: 2341391

Jolivet et al (1983) The pharmacology and clinical use of methotr. N.Engl.J.Med. 309 1094 PMID: 6353235

Winter-Vann et al (2003) Targeting Ras signaling through inhibition of carboxyl methylation: an unexpected property of methotr. Proc.Natl.Acad.Sci.U.S.A. 100 6529 PMID: 12750467

If you know of a relevant reference for Methotrexate, please let us know.

Keywords: Methotrexate, Methotrexate supplier, Cytotoxic, agent, Dihydrofolate, reductases, inhibitors, inhibits, Cell, Cycle, DHFR, chemotherapeutics, Reductase, Inhibitors, 1230, Tocris Bioscience

⚠ WARNING: This product can expose you to chemicals including Methotrexate, which is known to the State of California to cause reproductive toxicity with developmental effects. For more information, go to

2 Citations for Methotrexate

Citations are publications that use Tocris products. Selected citations for Methotrexate include:

Robinson et al (2013) RB1 status in triple negative breast cancer cells dictates response to radiation treatment and selective therapeutic drugs. PLoS One 8 e78641 PMID: 24265703

Kumar et al (2018) Comparison of cell-based assays to quantify treatment effects of anticancer drugs identifies a new application for Bodipy-L-cystine to measure apoptosis. Sci Rep 8 16363 PMID: 30397244

Do you know of a great paper that uses Methotrexate from Tocris? Please let us know.

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Literature in this Area

Tocris offers the following scientific literature in this area to showcase our products. We invite you to request* your copy today!

*Please note that Tocris will only send literature to established scientific business / institute addresses.

Cell Cycle and DNA Damage Research Product Guide

Cell Cycle and DNA Damage Research Product Guide

This product guide provides a review of the cell cycle and DNA damage research area and lists over 150 products, including research tools for:

  • Cell Cycle and Mitosis
  • DNA Damage Repair
  • Targeted Protein Degradation
  • Ubiquitin Proteasome Pathway
  • Chemotherapy Targets
Cell Cycle & DNA Damage Repair Poster

Cell Cycle & DNA Damage Repair Poster

In normal cells, each stage of the cell cycle is tightly regulated, however in cancer cells many genes and proteins that are involved in the regulation of the cell cycle are mutated or over expressed. This poster summarizes the stages of the cell cycle and DNA repair. It also highlights strategies for enhancing replicative stress in cancer cells to force mitotic catastrophe and cell death.