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
USD 970 In stock
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2 Customer Reviews

  • A. Viability comparison of REH vector control, BCL6 overexpression, or BCL6 overexpression cells pre-treated with 79-6 (125μM) following exposure to three chemotherapy drugs (Ara-C [1 μM], MTX [50 μM], VCR [25 μM]). (* = p < 0.05 BCL6 OX to vector control and # = p < 0.05 BCL6 OX to BCL6 + 79-6).

    Oncotarget, 2016, 7(17):23439-53. Methotrexate purchased from Selleck.

    Toxicity of MTX (3) and toxic analogues 5 a-e on a) hY1R-expressing MDA-MB-468 and b) non-hYR-expressing HEK293 cells. Cell viability was determined by resazurin assay. Bars represent the mean±SEM of at least three independent experiments performed in triplicate. Measurements were normalized by using only HBSS, pH 7, treated cells (set at 100 %) and ethanol-treated cells (set at 0 %). Statistical significances refer to only HBSS, pH 7, treated cells indicated as a dashed line.

    ChemMedChem, 2015, 10(5):804-14.. Methotrexate purchased from Selleck.

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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 MX7jfZRwfG:6aXPpeJkh[XO|YYm= MWTJR|UxRTRwODDuUS=> MlnIPFY{OjRzMx?=
L cells NX\EV5dj[3m2b4TvfIlkcXS7IHHzd4F6 NIjFbZJKSzVyPUiuN{BvVQ>? MmDIPFY{OjRzMx?=
D54 NHjpWXFkgXSxdH;4bYNqfHliYYPzZZk> M1XyUmlEPTB;MU[gcm0> NVrBe3RRQDZ|MkSxNy=>
143B(TK-) NXLpeldO[3m2b4TvfIlkcXS7IHHzd4F6 M130c2lEPTB;OD64JI5O NVO3UHhxQDZ|MkSxNy=>
A549 M33rTIN6fG:2b4jpZ4l1gSCjc4PhfS=> NXfle4xiUUN3ME2zNUBvVQ>? MknmPFY{OjRzMx?=
H460 Mk\ZZ5l1d3SxeHnjbZR6KGG|c3H5 NXXkUoM6UUN3ME25MlUhdk1? MlzJPFY{OjRzMx?=
Daoy NGO1WZVkgXSxdH;4bYNqfHliYYPzZZk> NGX0cmVKSzVyPUmgcm0> M1XvRVg3OzJ2MUO=
U373MG MVXjfZRwfG:6aXPpeJkh[XO|YYm= MWjJR|UxRTF{IH7N M3\hS|g3OzJ2MUO=
Vero NW\aOWlC[3m2b4TvfIlkcXS7IHHzd4F6 MnvlTWM2OD17LkKgcm0> MWO4OlMzPDF|
SCC25  Mlr1S5Jwf3SqIHnubIljcXSxcomgZZN{[Xl? MVf+NUDPxE1? NU\rXHNPUUN3ME2yO{BvVQ>? NH3KU486ODJ{N{m1
NCI-H460 M2faNGdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 NHy2S5N,OSEQvF2= NITwPI1KSzVyPUK4JI5O MVi5NFIzPzl3
NCI-H522 M4fNfGdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 MXH+NUDPxE1? MYfJR|UxRTJ{OTDuUS=> MWK5NFIzPzl3
EKVX MoXVS5Jwf3SqIHnubIljcXSxcomgZZN{[Xl? M{mwfJ4yKM7:TR?= NWT4N3Q4UUN3ME6xNFAxKG6P NFj5Wmo6ODJ{N{m1
HCT-116 M3X5Rmdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 Ml7zglEh|ryP NFGzTmtKSzVyPUOwJI5O NInBc|Y6ODJ{N{m1
HCT-15 MkPqS5Jwf3SqIHnubIljcXSxcomgZZN{[Xl? MmjnglEh|ryP NG\NXodKSzVyPUOwJI5O NGf4cmo6ODJ{N{m1
HT29 M2[2bGdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 MXP+NUDPxE1? MnL0TWM2OD1|MjDuUS=> MlLoPVAzOjd7NR?=
SW-620 NV7zOYpDT3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= M2nqbJ4yKM7:TR?= MkOzTWM2OD1|MzDuUS=> NVPyc5c{QTB{Mke5OS=>
KM12 NGjLPYRIem:5dHigbY5pcWKrdH;yfUBie3OjeR?= MWT+NUDPxE1? M3fofWlEPTB;NEKgcm0> NWewPXh[QTB{Mke5OS=>
SF-539 MWDHdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? MoLxglEh|ryP MmPKTWM2OD1|NTDuUS=> M3\sfFkxOjJ5OUW=
SF-268 Mn7tS5Jwf3SqIHnubIljcXSxcomgZZN{[Xl? NXrYZoJ[hjFizszN NUPlTWluUUN3ME21NkBvVQ>? NFH5fGY6ODJ{N{m1
SNB-75 MlvGS5Jwf3SqIHnubIljcXSxcomgZZN{[Xl? MVL+NUDPxE1? MWTJR|UxRjFyMECwJI5O M4CwNlkxOjJ5OUW=
LOX IMVI MVHHdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? NGLsV4x,OSEQvF2= NI[wRm1KSzVyPUK2JI5O Ml;IPVAzOjd7NR?=
UACC-62 NITCepBIem:5dHigbY5pcWKrdH;yfUBie3OjeR?= NYK4U|ZKhjFizszN MlfqTWM2OD1{ODDuUS=> NHfC[pg6ODJ{N{m1
SK-MEL-5 M{PQ[mdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 M2PFVp4yKM7:TR?= NYLZXo1[UUN3ME24O{BvVQ>? Mo\iPVAzOjd7NR?=
MALME-3M M13RNGdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 MXT+NUDPxE1? MoP0TWM2OD5zMECwJI5O MUG5NFIzPzl3
SK-MEL-28 MXPHdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? MU\+NUDPxE1? MoK5TWM2OD5zMECwJI5O M{HVUlkxOjJ5OUW=
OVCAR-5 MWPHdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? MnXvglEh|ryP MofNTWM2OD5zMECwJI5O Mn20PVAzOjd7NR?=
OVCAR-3 NV;qcop1T3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= Ml;DglEh|ryP NX[wR5JsUUN3ME2zPVghdk1? NEC3PFg6ODJ{N{m1
786-0 Mm\jS5Jwf3SqIHnubIljcXSxcomgZZN{[Xl? NYrQc3d3hjFizszN NXTvTHkyUUN3ME2zN{BvVQ>? Mn;uPVAzOjd7NR?=
UO-31 M3LBOWdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 MoKzglEh|ryP M2rvSWlEPTB;MUmxJI5O M4DlWFkxOjJ5OUW=
ACHN NXfIN|h5T3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= MlXTglEh|ryP M{fSXWlEPTB;NECgcm0> MlezPVAzOjd7NR?=
MCF7 MWHHdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? M3jKdJ4yKM7:TR?= M4fVUWlEPTB;M{[gcm0> M4TBVVkxOjJ5OUW=
MCF7-ADR NYT0U4o1T3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= MlfRglEh|ryP MkLyTWM2OD15ODDuUS=> MkLuPVAzOjd7NR?=
PC-3 NGnUVI9Iem:5dHigbY5pcWKrdH;yfUBie3OjeR?= NGDkZod,OSEQvF2= Mnf6TWM2OD1{IH7N NH;nUXU6ODJ{N{m1
DU-145 MWrHdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? MWL+NUDPxE1? Mk\JTWM2OD1{MzDuUS=> NHLITGw6ODJ{N{m1
CCRF-CEM NH[yVYlIem:5dHigbY5pcWKrdH;yfUBie3OjeR?= MXzFR|UxRTF2LkSgcm0> MX2xNFk2PjJ{MR?=
R1 MYnHdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? MWfFR|UxRTZ5NTDuUS=> NU\xZ49tOTB7NU[yNlE>
R2(Bos) Mo[1S5Jwf3SqIHnubIljcXSxcomgZZN{[Xl? MnvmSWM2OD1zNkCwJI5O MlqyNVA6PTZ{MkG=
FaDu MXjHdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? MXTFR|UxRTFzLkOgcm0> MUOxNFk2PjJ{MR?=
A253 NX;hUG4xT3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= NV7vVlExTUN3ME2xOE42KG6P M4f5PFExQTV4MkKx
HT-29 Mm\4Z5l1d3SxeHnjbZR6KGG|c3H5 MnfhTWM2OD1|LkS1JO69VQ>? NFzYXnkzOzl4OEiyOC=>
COLO-320 DM M37vPIN6fG:2b4jpZ4l1gSCjc4PhfS=> NGO0dmZKSzVyPUWuNlUh|ryP M1jLR|I{QTZ6OEK0
COLO 205 MVvjfZRwfG:6aXPpeJkh[XO|YYm= M3HmPWlEPTB;Mz6yO{DPxE1? MnvkNlM6Pjh6MkS=
BGC-823 M2jRN4N6fG:2b4jpZ4l1gSCjc4PhfS=> M4HQeWlEPTB;MD6xNUDPxE1? MnfQNVg2PTV3NkK=
Hela NWDpb3h5[3m2b4TvfIlkcXS7IHHzd4F6 MVjJR|UxRTBwMTFOwG0> MVexPFU2PTV4Mh?=
Bel-7402 M3HWeYN6fG:2b4jpZ4l1gSCjc4PhfS=> NYT3S4htUUN3ME24O{46KM7:TR?= NHTi[YIyQDV3NUW2Ni=>

... Click to View More Cell Line Experimental Data

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:
2% DMSO+30% PEG 300+5% Tween 80+ddH2O

* 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
Synonyms NCI-C04671

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Clinical Trial Information

NCT Number Recruitment Conditions Sponsor/Collaborators Start Date Phases
NCT01338987 Active, not recruiting Myelodysplastic Syndrome RAEB 2|Acute Lymphocytic Leukemia|Acute Myelogenous Leukemia|Myelodysplastic Syndrome RAEB 1 National Cancer Institute (NCI)|National Institutes of Health Clinical Center (CC) March 30, 2011 Phase 2
NCT02250937 Recruiting Acute Myeloid Leukemia|Myelodysplastic Syndrome M.D. Anderson Cancer Center October 27, 2014 Phase 2
NCT01875237 Active, not recruiting Leukemia|Myeloma|Myeloproliferative Diseases M.D. Anderson Cancer Center|Bellicum Pharmaceuticals December 27, 2013 Phase 1|Phase 2
NCT02385110 Recruiting Leukemia M.D. Anderson Cancer Center September 23, 2015 Phase 2
NCT03007147 Not yet recruiting B Acute Lymphoblastic Leukemia With t(9;22)(q34.1;q11.2); BCR-ABL1|BCR-ABL1 Fusion Protein Expression|Minimal Residual Disease|Philadelphia Chromosome Positive|T Acute Lymphoblastic Leukemia|Untreated Adult Acute Lymphoblastic Leukemia|Untreated Childhood Acute Lymphoblastic Leukemia Childrens Oncology Group|National Cancer Institute (NCI) July 2017 Phase 3
NCT02724904 Not yet recruiting Lymphoma Massachusetts General Hospital|Adienne SA|Dana-Farber Cancer Institute May 2017 --

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID