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 NWmyO5FU[3m2b4TvfIlkcXS7IHHzd4F6 NIXnUZRKSzVyPUSuPEBvVQ>? Ml\rPFY{OjRzMx?=
L cells NGHaeG9kgXSxdH;4bYNqfHliYYPzZZk> NWflS|JWUUN3ME24MlMhdk1? M1LobVg3OzJ2MUO=
A549 M1vydoN6fG:2b4jpZ4l1gSCjc4PhfS=> MmLGTWM2OD1|MTDuUS=> NFThdm45PjN{NEGz
H460 MkLiZ5l1d3SxeHnjbZR6KGG|c3H5 NYTEeGFxUUN3ME25MlUhdk1? NGrMbIQ5PjN{NEGz
Daoy MW\jfZRwfG:6aXPpeJkh[XO|YYm= M4H6TmlEPTB;OTDuUS=> MWm4OlMzPDF|
U373MG M1O1ZoN6fG:2b4jpZ4l1gSCjc4PhfS=> MYXJR|UxRTF{IH7N MUG4OlMzPDF|
Vero Ml3DZ5l1d3SxeHnjbZR6KGG|c3H5 NFLtWXJKSzVyPUmuNkBvVQ>? NUK3bJJuQDZ|MkSxNy=>
SCC25  MVPHdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? M1;PUZ4yKM7:TR?= NGS0b5BKSzVyPUK3JI5O NFna[mI6ODJ{N{m1
NCI-H460 MXnHdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? Mn3GglEh|ryP NXLFZ5NRUUN3ME2yPEBvVQ>? M2eyZ|kxOjJ5OUW=
NCI-H23 NHf5OVZIem:5dHigbY5pcWKrdH;yfUBie3OjeR?= NXnIfZJjhjFizszN NXvXelRKUUN3ME20N{BvVQ>? M{DO[|kxOjJ5OUW=
NCI-H522 MV7Hdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? M1iyd54yKM7:TR?= M{DrbmlEPTB;MkK5JI5O NXLHdFQ3QTB{Mke5OS=>
HCT-116 NXzMdVVqT3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= MmTIglEh|ryP NGnlW5BKSzVyPUOwJI5O NXrOcJlLQTB{Mke5OS=>
HCT-15 NYjNUWxIT3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= NUnYNZBqhjFizszN M1PtN2lEPTB;M{Cgcm0> NEXsbWk6ODJ{N{m1
HT29 MnnmS5Jwf3SqIHnubIljcXSxcomgZZN{[Xl? NWLy[FFjhjFizszN MULJR|UxRTN{IH7N MkHZPVAzOjd7NR?=
SW-620 MWPHdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? MXz+NUDPxE1? NFmyR3FKSzVyPUOzJI5O MWm5NFIzPzl3
KM12 NGPZOHJIem:5dHigbY5pcWKrdH;yfUBie3OjeR?= NUjDR5U4hjFizszN MULJR|UxRTR{IH7N MX:5NFIzPzl3
SF-539 MYTHdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? MkDFglEh|ryP NGHHO45KSzVyPUO1JI5O NFXxO5Q6ODJ{N{m1
SF-268 MlXTS5Jwf3SqIHnubIljcXSxcomgZZN{[Xl? M2jyXJ4yKM7:TR?= M365WWlEPTB;NUKgcm0> M{DOR|kxOjJ5OUW=
LOX IMVI NIP1dotIem:5dHigbY5pcWKrdH;yfUBie3OjeR?= MmO5glEh|ryP MnrtTWM2OD1{NjDuUS=> M4DS[VkxOjJ5OUW=
UACC-62 M3:zeWdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 MUT+NUDPxE1? NVfU[2VxUUN3ME2yPEBvVQ>? MXK5NFIzPzl3
SK-MEL-5 NYXNcHNIT3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= MlfmglEh|ryP MnnHTWM2OD16NzDuUS=> NFG3dpE6ODJ{N{m1
MALME-3M MYLHdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? NXKzOIFWhjFizszN M3i5N2lEPTB-MUCwNEBvVQ>? NV62NnY3QTB{Mke5OS=>
SK-MEL-28 Mo\aS5Jwf3SqIHnubIljcXSxcomgZZN{[Xl? MmL1glEh|ryP MYTJR|UxRjFyMECgcm0> MoS4PVAzOjd7NR?=
OVCAR-8 Mn2wS5Jwf3SqIHnubIljcXSxcomgZZN{[Xl? NIqzPXd,OSEQvF2= MVnJR|UxRTNzIH7N NX;aS4l6QTB{Mke5OS=>
OVCAR-5 NVmyfG55T3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= MXv+NUDPxE1? MX\JR|UxRjFyMECgcm0> Mm\wPVAzOjd7NR?=
OVCAR-3 NUPyOoVQT3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= M4fLRp4yKM7:TR?= NI\hd|JKSzVyPUO5PEBvVQ>? MUi5NFIzPzl3
786-0 M2j1[Gdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 NXzvcWl4hjFizszN M4qwemlEPTB;M{Ogcm0> NU[2ZopsQTB{Mke5OS=>
UO-31 NWLvR25ZT3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= M2LSR54yKM7:TR?= M33QfGlEPTB;MUmxJI5O NXnOXog6QTB{Mke5OS=>
ACHN M1;yZWdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 MWr+NUDPxE1? M2nCWWlEPTB;NECgcm0> MVS5NFIzPzl3
MCF7 NVXEdXB6T3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= NYL2O2dqhjFizszN M1i0NGlEPTB;M{[gcm0> M4TPU|kxOjJ5OUW=
MCF7-ADR NW\XUnA{T3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= M4DMeJ4yKM7:TR?= MmS5TWM2OD15ODDuUS=> NGq4bo06ODJ{N{m1
PC-3 NHXVXHVIem:5dHigbY5pcWKrdH;yfUBie3OjeR?= MX3+NUDPxE1? M1HtUWlEPTB;MjDuUS=> NHfFVZc6ODJ{N{m1
DU-145 NHLPUI9Iem:5dHigbY5pcWKrdH;yfUBie3OjeR?= NXrudmhQhjFizszN M2fXdGlEPTB;MkOgcm0> NVn3Umh7QTB{Mke5OS=>
R2(Bos) NEmxd2pIem:5dHigbY5pcWKrdH;yfUBie3OjeR?= NIPR[nBGSzVyPUG2NFAhdk1? NIPDRYUyODl3NkKyNS=>
R30dm M{PLN2dzd3e2aDDpcohq[mm2b4L5JIF{e2G7 M2Cwb2VEPTB;MUSgcm0> M4WzPVExQTV4MkKx
FaDu MYfHdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? NEPiNmFGSzVyPUGxMlMhdk1? MXSxNFk2PjJ{MR?=
A253 NV3mclRzT3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= MlnNSWM2OD1zND61JI5O M{Sze|ExQTV4MkKx
COLO-320 DM MUXjfZRwfG:6aXPpeJkh[XO|YYm= NIfXOmJKSzVyPUWuNlUh|ryP MVeyN|k3QDh{NB?=
COLO 205 Mlv0Z5l1d3SxeHnjbZR6KGG|c3H5 NET5c2RKSzVyPUOuNlch|ryP MljVNlM6Pjh6MkS=
BGC-823 NVPTbI0y[3m2b4TvfIlkcXS7IHHzd4F6 Mn2zTWM2OD1yLkGxJO69VQ>? M1HkVVE5PTV3NU[y
Hela M4fHR4N6fG:2b4jpZ4l1gSCjc4PhfS=> M4TQZWlEPTB;MD6xJO69VQ>? M1;OZVE5PTV3NU[y
Bel-7402 NU\2dnpI[3m2b4TvfIlkcXS7IHHzd4F6 NF20fWZKSzVyPUi3Mlkh|ryP MYqxPFU2PTV4Mh?=

... 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
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

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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