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= NHPPb5dKSzVyPUSuPEBvVQ>? MnnLPFY{OjRzMx?=
L cells M3zyOYN6fG:2b4jpZ4l1gSCjc4PhfS=> MV\JR|UxRThwMzDuUS=> M2TGTVg3OzJ2MUO=
D54 NYnjdnlE[3m2b4TvfIlkcXS7IHHzd4F6 NX:zO2UyUUN3ME2xOkBvVQ>? M{SyUlg3OzJ2MUO=
143B(TK-) MXzjfZRwfG:6aXPpeJkh[XO|YYm= NW\yXHVKUUN3ME24Mlghdk1? M4XRU|g3OzJ2MUO=
A549 NFrFb2hkgXSxdH;4bYNqfHliYYPzZZk> MY\JR|UxRTNzIH7N MkDUPFY{OjRzMx?=
H460 NVHte4pt[3m2b4TvfIlkcXS7IHHzd4F6 MVrJR|UxRTlwNTDuUS=> NGr5e5U5PjN{NEGz
Daoy MV;jfZRwfG:6aXPpeJkh[XO|YYm= M3rYbGlEPTB;OTDuUS=> M13XR|g3OzJ2MUO=
U373MG Mm\NZ5l1d3SxeHnjbZR6KGG|c3H5 M33wNGlEPTB;MUKgcm0> MX64OlMzPDF|
Vero M1e3WIN6fG:2b4jpZ4l1gSCjc4PhfS=> MoLOTWM2OD17LkKgcm0> NYH6b2pxQDZ|MkSxNy=>
SCC25  Mmi1S5Jwf3SqIHnubIljcXSxcomgZZN{[Xl? MYj+NUDPxE1? NWX6fnhjUUN3ME2yO{BvVQ>? NFfyVZo6ODJ{N{m1
NCI-H460 MoLVS5Jwf3SqIHnubIljcXSxcomgZZN{[Xl? M3XIRp4yKM7:TR?= MojETWM2OD1{ODDuUS=> NHzOc406ODJ{N{m1
NCI-H23 Mn\FS5Jwf3SqIHnubIljcXSxcomgZZN{[Xl? MnjGglEh|ryP M2r0SGlEPTB;NEOgcm0> MXO5NFIzPzl3
NCI-H522 NFrpV3RIem:5dHigbY5pcWKrdH;yfUBie3OjeR?= MUn+NUDPxE1? NV;ROIJwUUN3ME2yNlkhdk1? MlXLPVAzOjd7NR?=
EKVX NXfNVoRGT3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= M16xPJ4yKM7:TR?= MVnJR|UxRjFyMECgcm0> NUXB[WVbQTB{Mke5OS=>
HCT-116 MWHHdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? NEDRNoZ,OSEQvF2= MoLFTWM2OD1|MDDuUS=> M{i5elkxOjJ5OUW=
HT29 MW\Hdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? NWfYPG5VhjFizszN NYXRPWJHUUN3ME2zNkBvVQ>? MVy5NFIzPzl3
SW-620 MVnHdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? MlTwglEh|ryP MoPSTWM2OD1|MzDuUS=> MVe5NFIzPzl3
KM12 MVXHdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? M4fxNZ4yKM7:TR?= MmrjTWM2OD12MjDuUS=> MYK5NFIzPzl3
SF-539 M4XEO2dzd3e2aDDpcohq[mm2b4L5JIF{e2G7 MWL+NUDPxE1? M4qxZmlEPTB;M{Wgcm0> MkO5PVAzOjd7NR?=
SF-268 NInMVHBIem:5dHigbY5pcWKrdH;yfUBie3OjeR?= MnfvglEh|ryP NXvCW2hJUUN3ME21NkBvVQ>? M1PVTFkxOjJ5OUW=
SNB-75 M{O3OGdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 MnvBglEh|ryP NFPS[FJKSzVyPkGwNFAxKG6P NF;zUpE6ODJ{N{m1
LOX IMVI NFLYb4ZIem:5dHigbY5pcWKrdH;yfUBie3OjeR?= NGnzWVR,OSEQvF2= M3PwNWlEPTB;Mk[gcm0> MkXlPVAzOjd7NR?=
UACC-62 MW\Hdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? M4fhT54yKM7:TR?= MkPrTWM2OD1{ODDuUS=> NFHNO|Q6ODJ{N{m1
SK-MEL-5 M2O2WGdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 NGTGOlN,OSEQvF2= M1vVSmlEPTB;OEegcm0> NFH5fFA6ODJ{N{m1
MALME-3M MY\Hdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? NUC5cld4hjFizszN M2frNGlEPTB-MUCwNEBvVQ>? M4eyOVkxOjJ5OUW=
SK-MEL-28 MmrvS5Jwf3SqIHnubIljcXSxcomgZZN{[Xl? MUP+NUDPxE1? NV30d4JnUUN3ME6xNFAxKG6P MYK5NFIzPzl3
OVCAR-5 M3TZV2dzd3e2aDDpcohq[mm2b4L5JIF{e2G7 Mk\YglEh|ryP Mn76TWM2OD5zMECwJI5O NHXHcGw6ODJ{N{m1
OVCAR-3 NVPJRYt1T3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= NET0XHZ,OSEQvF2= M{\nfmlEPTB;M{m4JI5O M3nKe|kxOjJ5OUW=
786-0 MonjS5Jwf3SqIHnubIljcXSxcomgZZN{[Xl? MW\+NUDPxE1? NYnlTWZvUUN3ME2zN{BvVQ>? M3jnflkxOjJ5OUW=
UO-31 NVTDTphMT3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= MnG4glEh|ryP MojSTWM2OD1zOUGgcm0> NVLRfI5ZQTB{Mke5OS=>
ACHN MmXaS5Jwf3SqIHnubIljcXSxcomgZZN{[Xl? MVv+NUDPxE1? M1K0PGlEPTB;NECgcm0> MWW5NFIzPzl3
MCF7-ADR M3PRWWdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 MlrhglEh|ryP M3TKeWlEPTB;N{igcm0> MV:5NFIzPzl3
PC-3 NUj1U457T3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= NGDXUXZ,OSEQvF2= MXLJR|UxRTJibl2= MlnuPVAzOjd7NR?=
DU-145 NUHvcIs3T3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= MUP+NUDPxE1? MmrETWM2OD1{MzDuUS=> MXG5NFIzPzl3
CCRF-CEM M4joWmdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 M3LCOmVEPTB;MUSuOEBvVQ>? M4\rR|ExQTV4MkKx
R1 MnjMS5Jwf3SqIHnubIljcXSxcomgZZN{[Xl? NVPuRYlsTUN3ME22O|Uhdk1? MW[xNFk2PjJ{MR?=
R2(Bos) MW\Hdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? NYP3cXZtTUN3ME2xOlAxKG6P MVWxNFk2PjJ{MR?=
R30dm NV;nU5dlT3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= M1y0UGVEPTB;MUSgcm0> NXXrN5BOOTB7NU[yNlE>
FaDu NVHqb4g1T3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= NWnKRmxOTUN3ME2xNU4{KG6P M1jzeVExQTV4MkKx
A253 M4PZc2dzd3e2aDDpcohq[mm2b4L5JIF{e2G7 NF;mfXVGSzVyPUG0MlUhdk1? NUP0O|Q4OTB7NU[yNlE>
HT-29 NYO5[VNY[3m2b4TvfIlkcXS7IHHzd4F6 NXjRSnRlUUN3ME2zMlQ2KM7:TR?= NVm3bHFxOjN7Nki4NlQ>
COLO-320 DM M3zlS4N6fG:2b4jpZ4l1gSCjc4PhfS=> NHTkZndKSzVyPUWuNlUh|ryP NVLZdGlkOjN7Nki4NlQ>
COLO 205 NYWydYlv[3m2b4TvfIlkcXS7IHHzd4F6 MWrJR|UxRTNwMkeg{txO NH\PPWIzOzl4OEiyOC=>
BGC-823 MmXyZ5l1d3SxeHnjbZR6KGG|c3H5 MVLJR|UxRTBwMUGg{txO NF7iUpAyQDV3NUW2Ni=>
Bel-7402 NVy2OJJJ[3m2b4TvfIlkcXS7IHHzd4F6 NYnKVndEUUN3ME24O{46KM7:TR?= Mni5NVg2PTV3NkK=

... 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
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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DHFR Signaling Pathway Map

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID