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  4. Methotrexate hydrate

Methotrexate hydrate  (Synonyms: Amethopterin hydrate; CL14377 hydrate; WR19039 hydrate)

Cat. No.: HY-14519B Purity: 99.57%
COA
SDS
Handling Instructions Technical Support

Methotrexate hydrate is a substance that affects cell growth and division. It is commonly used to fight certain diseases, especially cancer and autoimmune diseases. It works by inhibiting the process by which cells synthesize DNA.

For research use only. We do not sell to patients.

CAS No. : 133073-73-1

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Customer Review

Based on 76 publication(s) in Google Scholar

Other Forms of Methotrexate hydrate:

Methotrexate hydrate Related Antibodies

Top Publications Citing Use of Products

76 Publications Citing Use of MCE Methotrexate hydrate

Cell Imaging/Staining
Cell Proliferation/Viability Assay
RT-PCR
Cell Migration/Invasion Assay

    Methotrexate hydrate purchased from MedChemExpress. Usage Cited in: Nat Commun. 2025 Feb 28;16(1):2071.  [Abstract]

    OIS cells were infected with shCtrl orshACSS2 followed by selection and treated with or without MTX (40 μM) for 2 days. Cells were stained with γH2AX.

    Methotrexate hydrate purchased from MedChemExpress. Usage Cited in: Nat Commun. 2025 Feb 28;16(1):2071.  [Abstract]

    OIS cells were infected with shCtrl orshACSS2 followed by selection and treated with or without MTX (40 μM) for 2 days. The expression of SASP genes was analysed by qRT–PCR.

    Methotrexate hydrate purchased from MedChemExpress. Usage Cited in: J Adv Res. 2025 Aug:74:609-620.  [Abstract]

    The cell viability of MH7A cells treated with methotrexate (1 μM) for 24 h was determined by the CCK-8 method.

    Methotrexate hydrate purchased from MedChemExpress. Usage Cited in: J Adv Res. 2025 Aug:74:609-620.  [Abstract]

    The cell migration of MH7A cells treated with methotrexate (1 μM) for 24 h.

    Methotrexate hydrate purchased from MedChemExpress. Usage Cited in: J Clin Invest. 2023 Jul 3;133(13):e169993.  [Abstract]

    A CellTiter-Glo® luminescent cell viability assay was performed on CAL51 WT and MLL4 KO cells, which were treated with 0, 0.04, or 0.4 μM Methotrexate (MTX) in the presence of H2O, 50 μM thymidine, or 50 μM inosine.

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    Description

    Methotrexate hydrate is a substance that affects cell growth and division. It is commonly used to fight certain diseases, especially cancer and autoimmune diseases. It works by inhibiting the process by which cells synthesize DNA.

    In Vitro

    Methotrexate (hydrate) is a biochemical reagent that can be used as a biological material or organic compound for life science related research.

    MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

    Methotrexate hydrate Related Antibodies
    Formula

    C20H22N8O5.xH2O
    CAS No.
    Appearance

    Solid

    Color

    Yellow to orange

    SMILES

    NC1=NC(N)=C2C(N=CC(CN(C)C3=CC=C(C(N[C@@H](CCC(O)=O)C(O)=O)=O)C=C3)=N2)=N1.O.[x]
    Shipping

    Room temperature in continental US; may vary elsewhere.
    Storage

    4°C, sealed storage, away from moisture and light

    *In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)

    Solvent & Solubility
    In Vitro: 

    DMSO : 10 mg/mL (ultrasonic and warming and heat to 60°C; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

    • Molarity Calculator

    • Dilution Calculator

    Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

    Mass
    =
    Concentration
    ×
    Volume
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    Molecular Weight *

    Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

    This equation is commonly abbreviated as: C1V1 = C2V2

    Concentration (start)

    C1

    ×
    Volume (start)

    V1

    =
    Concentration (final)

    C2

    ×
    Volume (final)

    V2

    In Vivo:

    Select the appropriate dissolution method based on your experimental animal and administration route.

    For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
    To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
    The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

    • Protocol 1

      Add each solvent one by one:  10% DMSO    40% PEG300    5% Tween-80    45% Saline

      Solubility: ≥ 1 mg/mL; Clear solution

      This protocol yields a clear solution of ≥ 1 mg/mL (saturation unknown).

      Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (10.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.

      Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
    • Protocol 2

      Add each solvent one by one:  10% DMSO    90% (20% SBE-β-CD in Saline)

      Solubility: ≥ 1 mg/mL; Clear solution

      This protocol yields a clear solution of ≥ 1 mg/mL (saturation unknown).

      Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (10.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.

      Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
    In Vivo Dissolution Calculator
    Please enter the basic information of animal experiments:

    Dosage

    mg/kg

    Animal weight
    (per animal)

    g

    Dosing volume
    (per animal)

    μL

    Number of animals

    Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
    Please enter your animal formula composition:
    %
    DMSO +
    % 
    +
    %
    Tween-80 +
    %
    Saline
    Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
    The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
    Calculation results:
    Working solution concentration: mg/mL
    Method for preparing stock solution: mg drug dissolved in μL  DMSO (Stock solution concentration: mg/mL).

    *In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)

    The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only. If necessary, please contact MedChemExpress (MCE).
    Method for preparing in vivo working solution for animal experiments: Take μL DMSO stock solution, add μL . μL , mix evenly, next add μL Tween 80, mix evenly, then add μL Saline.
     If the continuous dosing period exceeds half a month, please choose this protocol carefully.
    Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
    Purity & Documentation

    Purity: 99.57%

    SDS (644 KB)

    COA (248 KB) HNMR (134 KB) RP-HPLC (225 KB) LCMS (95 KB)

    Handling Instructions (2659 KB)
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    Methotrexate hydrate Related Classifications

    Help & FAQs
    • Do most proteins show cross-species activity?

      Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

    Keywords:

    Methotrexate133073-73-1AmethopterinCL14377WR19039CL 14377CL-14377WR 19039WR-19039Biochemical Assay ReagentsAntibioticBacterialbiochemical assayreagentInhibitorinhibitorinhibit

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