Catalog No.S1210 Synonyms: NCI-C04671

Methotrexate Chemical Structure

Molecular Weight(MW): 454.44

Methotrexate (MTX), analog of folic acid, is a nonspecific inhibitor of the dihydrofolate reductase(DHFR) of bacteria and cancerous cells as well as normal cells. It forms an inactive ternary complex with DHFR and NADPH.

Size Price Stock Quantity  
In DMSO USD 134 In stock
USD 97 In stock
USD 197 In stock
Bulk Discount

Free Overnight Delivery on orders over $ 500
Next day delivery by 10:00 a.m. Order now.

Cited by 22 Publications

Purity & Quality Control

Choose Selective DHFR Inhibitors

Biological Activity

Description Methotrexate (MTX), analog of folic acid, is a nonspecific inhibitor of the dihydrofolate reductase(DHFR) of bacteria and cancerous cells as well as normal cells. It forms an inactive ternary complex with DHFR and NADPH.
hDHFR [4]
(Activated peripheral T cells)
24 nM
In vitro

Methotrexate (0.1-10 mM) induces apoptosis of in vitro activated T cells from human peripheral blood. Methotrexate achieves clonal deletion of activated T cells in mixed lymphocyte reactions. Methotrexate can selectively delete activated peripheral blood T cells by a CD95-independent pathway. [1] Methotrexate is taken up by cells via the reduced folate carrier and then is converted within the cells to polyglutamates. Methotrexate leads to diminished production of leukotriene B4 by neutrophils stimulated ex vivo. Methotrexate polyglutamates inhibit the enzyme aminoimidazolecarboxamidoadenosineribonucleotide (AICAR) transformylase more potently than the other enzymes involved in purine biosynthesis. Methotrexate is also known to suppress TNF activity by suppressing TNF-induced nuclear factor-κB activation in vitro, in part related to a reduction in the degradation and inactivation of an inhibitor of this factor, IκBα, and probably related to the release of adenosine. Methotrexate suppresses the production of both TNF and IFN-γ by T-cell-receptor-primed T lymphocytes from both healthy human donors and RA patients. Methotrexate treatment is associated with a significant decrease of TNF-α-positive CD4+ T cells, while the number of T cells expressing the anti-inflammatory cytokine IL-10 increased. [2]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
P388D1 M3PnPIN6fG:2b4jpZ4l1gSCjc4PhfS=> NV\VUmk2UUN3ME20Mlghdk1? MoXVPFY{OjRzMx?=
L cells NUnrT3JG[3m2b4TvfIlkcXS7IHHzd4F6 M{n1XmlEPTB;OD6zJI5O NFTK[XY5PjN{NEGz
D54 NGrWfZRkgXSxdH;4bYNqfHliYYPzZZk> M1;3TWlEPTB;MU[gcm0> MoD1PFY{OjRzMx?=
143B(TK-) NGnq[oRkgXSxdH;4bYNqfHliYYPzZZk> MofqTWM2OD16Lkigcm0> NHTIcVM5PjN{NEGz
U87MG NUDKPVhS[3m2b4TvfIlkcXS7IHHzd4F6 M37lUmlEPTB;MkKgcm0> NGX1Znk5PjN{NEGz
A549 NYX5[FQ1[3m2b4TvfIlkcXS7IHHzd4F6 NVzERoU3UUN3ME2zNUBvVQ>? MVW4OlMzPDF|
H460 MWfjfZRwfG:6aXPpeJkh[XO|YYm= MkDLTWM2OD17LkWgcm0> NIfCdGY5PjN{NEGz
Daoy NW\GW3BW[3m2b4TvfIlkcXS7IHHzd4F6 M4exOmlEPTB;OTDuUS=> Mli2PFY{OjRzMx?=
Vero MYPjfZRwfG:6aXPpeJkh[XO|YYm= Mm\zTWM2OD17LkKgcm0> MWG4OlMzPDF|
SCC25  Mon1S5Jwf3SqIHnubIljcXSxcomgZZN{[Xl? NIXSSFB,OSEQvF2= M{n6dmlEPTB;Mkegcm0> NWjvSFFLQTB{Mke5OS=>
NCI-H460 M3zZOmdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 NFm0dnl,OSEQvF2= MVnJR|UxRTJ6IH7N Mmf2PVAzOjd7NR?=
NCI-H23 M{e4VWdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 MUL+NUDPxE1? Mnr3TWM2OD12MzDuUS=> NUXkSpRSQTB{Mke5OS=>
NCI-H522 NHnaeVFIem:5dHigbY5pcWKrdH;yfUBie3OjeR?= NVfWW2NXhjFizszN NULwNphrUUN3ME2yNlkhdk1? NEDhUYM6ODJ{N{m1
HCT-116 MoPPS5Jwf3SqIHnubIljcXSxcomgZZN{[Xl? MX\+NUDPxE1? NXvkNWN{UUN3ME2zNEBvVQ>? MnjoPVAzOjd7NR?=
HCT-15 NGP5UGRIem:5dHigbY5pcWKrdH;yfUBie3OjeR?= MnzrglEh|ryP M1ryVWlEPTB;M{Cgcm0> NGPIPGw6ODJ{N{m1
HT29 M{j0Wmdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 MoPMglEh|ryP M2O3UWlEPTB;M{Kgcm0> NV3YPGZHQTB{Mke5OS=>
SW-620 MkHmS5Jwf3SqIHnubIljcXSxcomgZZN{[Xl? M3LTSZ4yKM7:TR?= NUHIeHZ4UUN3ME2zN{BvVQ>? M3XFTlkxOjJ5OUW=
KM12 NG\KZ3FIem:5dHigbY5pcWKrdH;yfUBie3OjeR?= MUX+NUDPxE1? MonSTWM2OD12MjDuUS=> NE\CPWI6ODJ{N{m1
SF-539 MYXHdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? NFfrRoh,OSEQvF2= NHrWWXBKSzVyPUO1JI5O M3XsZVkxOjJ5OUW=
SF-268 M4\5RWdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 MVr+NUDPxE1? MkD6TWM2OD13MjDuUS=> Mn3oPVAzOjd7NR?=
SNB-75 MkP1S5Jwf3SqIHnubIljcXSxcomgZZN{[Xl? MUL+NUDPxE1? MortTWM2OD5zMECwNEBvVQ>? NVHDbFQyQTB{Mke5OS=>
LOX IMVI NGLhdZhIem:5dHigbY5pcWKrdH;yfUBie3OjeR?= NV3pOoJvhjFizszN M2T5U2lEPTB;Mk[gcm0> MlzNPVAzOjd7NR?=
UACC-62 M4\ve2dzd3e2aDDpcohq[mm2b4L5JIF{e2G7 NHjpVIx,OSEQvF2= NH;pT2lKSzVyPUK4JI5O NWfyOYF{QTB{Mke5OS=>
SK-MEL-5 NIH4RXBIem:5dHigbY5pcWKrdH;yfUBie3OjeR?= M{nodJ4yKM7:TR?= NGHlbnNKSzVyPUi3JI5O MXy5NFIzPzl3
SK-MEL-28 MY\Hdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? NHHGcHh,OSEQvF2= NHzsZ2RKSzVyPkGwNFAhdk1? MWe5NFIzPzl3
OVCAR-8 NVzxN4RFT3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= M3nr[54yKM7:TR?= M1zCZWlEPTB;M{Ggcm0> MUK5NFIzPzl3
OVCAR-5 Ml3lS5Jwf3SqIHnubIljcXSxcomgZZN{[Xl? MlfPglEh|ryP NGHjNZlKSzVyPkGwNFAhdk1? M4mxO|kxOjJ5OUW=
OVCAR-3 NUf1NXBUT3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= NYGycnkyhjFizszN MUfJR|UxRTN7ODDuUS=> M4myfFkxOjJ5OUW=
786-0 NF7vXGxIem:5dHigbY5pcWKrdH;yfUBie3OjeR?= NWHuXmZThjFizszN M{PXbWlEPTB;M{Ogcm0> NH7j[Jo6ODJ{N{m1
UO-31 M4H5[mdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 MlvvglEh|ryP NWfTOnpqUUN3ME2xPVEhdk1? NFTBbFM6ODJ{N{m1
ACHN NVPR[29lT3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= M3XpVJ4yKM7:TR?= M3npcWlEPTB;NECgcm0> MoniPVAzOjd7NR?=
MCF7 M{TaVmdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 NXXKell[hjFizszN NVzHeXRxUUN3ME2zOkBvVQ>? NE[yVlM6ODJ{N{m1
MCF7-ADR NHHoOJRIem:5dHigbY5pcWKrdH;yfUBie3OjeR?= NGnHS5B,OSEQvF2= M2fqWWlEPTB;N{igcm0> M2PPR|kxOjJ5OUW=
PC-3 MVHHdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? M1ftbJ4yKM7:TR?= M1TYfmlEPTB;MjDuUS=> NVvBeJc{QTB{Mke5OS=>
DU-145 MWfHdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? NEHqV5Z,OSEQvF2= MWHJR|UxRTJ|IH7N NIq4eHA6ODJ{N{m1
CCRF-CEM NUDYeG9XT3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= MV;FR|UxRTF2LkSgcm0> NH\qemcyODl3NkKyNS=>
R1 M1vPc2dzd3e2aDDpcohq[mm2b4L5JIF{e2G7 NGXvT|BGSzVyPU[3OUBvVQ>? MWCxNFk2PjJ{MR?=
R2(Bos) M1vlPWdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 MXnFR|UxRTF4MECgcm0> M2PqTFExQTV4MkKx
R30dm MV\Hdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? Mn3FSWM2OD1zNDDuUS=> M1fzelExQTV4MkKx
FaDu M4fjNWdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 MV;FR|UxRTFzLkOgcm0> M{fmcFExQTV4MkKx
A253 M1i5bGdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 NXXWU3ZKTUN3ME2xOE42KG6P MmTrNVA6PTZ{MkG=
HT-29 M13wfIN6fG:2b4jpZ4l1gSCjc4PhfS=> NFT3cIVKSzVyPUOuOFUh|ryP MmWzNlM6Pjh6MkS=
COLO-320 DM MmLnZ5l1d3SxeHnjbZR6KGG|c3H5 MXzJR|UxRTVwMkWg{txO NFrrVmczOzl4OEiyOC=>
COLO 205 NYjTNopO[3m2b4TvfIlkcXS7IHHzd4F6 M{SwWmlEPTB;Mz6yO{DPxE1? NITGXW0zOzl4OEiyOC=>
Hela M13SZ4N6fG:2b4jpZ4l1gSCjc4PhfS=> NVXZemFpUUN3ME2wMlEh|ryP MonHNVg2PTV3NkK=
Bel-7402 NX;2U5pt[3m2b4TvfIlkcXS7IHHzd4F6 MVnJR|UxRTh5Lkmg{txO MlfTNVg2PTV3NkK=

... Click to View More Cell Line Experimental Data

Methods Test Index PMID
Western blot
ppJAK1 / ppJAK2 / JAK1 / JAK2 ; 

PubMed: 26131691     

HDLM-2 cells treated with the indicated concentrations of methotrexate and aminopterin and blotted to show levels of total JAK1 (tJAK1), dual phosphorylated JAK1 (ppJAK1), total JAK2 (tJAK2), dual phosphorylated JAK2 (ppJAK2) and ß-actin shown as a loading control. Levels of ppJAK1 and ppJAK2 are reduced at high drug concentrations.

pSTAT1 / pSTAT3 / pSTAT5 / STAT1 / STAT3 / STAT5; 

PubMed: 26131691     

D) HDLM-2 cells treated with the indicated concentrations of methotrexate and aminopterin and blotted to show levels of total STAT1 (tSTAT1), phosphorylated STAT1 (pSTAT1), total STAT3 (tSTAT3), phosphorylated STAT3 (pSTAT3), total STAT5 (tSTAT5), phosphorylated STAT5 (pSTAT5) and β-actin shown as a loading control. Levels of pSTAT1 and pSTAT5 are reduced at most drug concentrations while levels of pSTAT3 appear to be unaffected.

AKT / p-AKT / mTOR / p-mTOR / Beclin 1 / HMGB1 ; 

PubMed: 26702616     

After the cells were exposed to 0.1 μM MTX for the indicated times, the cell lysates were subjected to Western blotting with specific antibodies. The results are representative of three independent experiments. β-actin was used as a loading control.

26131691 26702616
In vivo Methotrexate increases splenocyte AICAR content, raised adenosine concentrations in exudates from carrageenan-inflamed air pouches, and markedly inhibits leukocyte accumulation in inflamed air pouches in mice. Methotrexate-mediated reduction in leukocyte accumulation is partially reversed by injection of adenosine deaminase (ADA) into the air pouch, completely reverses by a specific adenosine A2 receptor antagonist, 3,7-dimethyl-1-propargylxanthine (DMPX), but not affected by an adenosine A1 receptor antagonist, 8-cyclopentyl-dipropylxanthine in mice. [3]


Solubility (25°C)

In vitro DMSO 90 mg/mL warmed (198.04 mM)
Water Insoluble
Ethanol Insoluble
In vivo Add solvents to the product individually and in order(Data is from Selleck tests instead of citations):
2% DMSO+30% PEG 300+5% Tween 80+ddH2O
For best results, use promptly after mixing.

* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.

Chemical Information

Molecular Weight 454.44


CAS No. 59-05-2
Storage powder
in solvent
Synonyms NCI-C04671

Bio Calculators

Molarity Calculator

Molarity Calculator

Calculate the mass, volume or concentration required for a solution. The Selleck molarity calculator is based on the following equation:

Mass (mg) = Concentration (mM) × Volume (mL) × Molecular Weight (g/mol)

  • Mass
    Molecular Weight

*When preparing stock solutions, please always use the batch-specific molecular weight of the product found on the via label and MSDS / COA (available on product pages).

Dilution Calculator

Dilution Calculator

Calculate the dilution required to prepare a stock solution. The Selleck dilution calculator is based on the following equation:

Concentration (start) x Volume (start) = Concentration (final) x Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2 ( Input Output )

  • C1

* When preparing stock solutions always use the batch-specific molecular weight of the product found on the vial label and MSDS / COA (available online).

The Serial Dilution Calculator Equation

  • Serial Dilutions

  • Computed Result

  • C1=C0/X C1: LOG(C1):
    C2=C1/X C2: LOG(C2):
    C3=C2/X C3: LOG(C3):
    C4=C3/X C4: LOG(C4):
    C5=C4/X C5: LOG(C5):
    C6=C5/X C6: LOG(C6):
    C7=C6/X C7: LOG(C7):
    C8=C7/X C8: LOG(C8):
Molecular Weight Calculator

Molecular Weight Calculator

Enter the chemical formula of a compound to calculate its molar mass and elemental composition:

Total Molecular Weight: g/mol

Tip: Chemical formula is case sensitive. C10H16N2O2 c10h16n2o2

Instructions to calculate molar mass (molecular weight) of a chemical compound:

To calculate molar mass of a chemical compound, please enter its chemical formula and click 'Calculate'.

Definitions of molecular mass, molecular weight, molar mass and molar weight:

Molecular mass (molecular weight) is the mass of one molecule of a substance and is expressed in the unified atomic mass units (u). (1 u is equal to 1/12 the mass of one atom of carbon-12)
Molar mass (molar weight) is the mass of one mole of a substance and is expressed in g/mol.

Molarity Calculator

Mass Concentration Volume Molecular Weight

Clinical Trial Information

NCT Number Recruitment interventions Conditions Sponsor/Collaborators Start Date Phases
NCT04109300 Not yet recruiting Genetic: HLADQA1*05A>G screening|Other: Standard of Care Inflammatory Bowel Diseases|Ulcerative Colitis|Crohn Disease Western University Canada September 1 2020 Not Applicable
NCT03642795 Not yet recruiting Other: Questionnaire Rheumatoid Polyarthritis Centre Hospitalier Universitaire de Nīmes January 2020 --
NCT03964259 Recruiting Drug: Intravenous fluids Lymphoma|Acute Lymphoblastic Leukemia|Pediatric Cancer|Pediatric ALL|Pediatric Lymphoma Virginia Commonwealth University October 2 2019 Phase 1
NCT03960177 Recruiting Drug: Glucarpidase Osteosarcoma OHSU Knight Cancer Institute|National Cancer Institute (NCI) March 27 2019 Early Phase 1

Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

Tel: +1-832-582-8158 Ext:3

If you have any other enquiries, please leave a message.

  • * Indicates a Required Field

DHFR Signaling Pathway Map

Related DHFR Products

Tags: buy Methotrexate | Methotrexate supplier | purchase Methotrexate | Methotrexate cost | Methotrexate manufacturer | order Methotrexate | Methotrexate distributor
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID