1. PI3K/Akt/mTOR
    Autophagy
    Metabolic Enzyme/Protease
  2. PI3K
    Autophagy
    Mitophagy
    Endogenous Metabolite

3-Methyladenine (Synonyms: 3-MA)

Cat. No.: HY-19312 Purity: 99.84%
Handling Instructions

3-Methyladenine is a potent inhibitor of autophagy and PI3K, blocking autophagy through its effect on PI3K, with IC50s of 25 μM and 60 μM for Vps34 and PI3Kγ, respectively.

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

3-Methyladenine Chemical Structure

3-Methyladenine Chemical Structure

CAS No. : 5142-23-4

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Free Sample (0.5-1 mg)   Apply now  
10 mM * 1 mL in DMSO USD 66 In-stock
Estimated Time of Arrival: December 31
50 mg USD 60 In-stock
Estimated Time of Arrival: December 31
100 mg USD 96 In-stock
Estimated Time of Arrival: December 31
200 mg USD 168 In-stock
Estimated Time of Arrival: December 31
500 mg USD 324 In-stock
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Customer Review

    3-Methyladenine purchased from MCE. Usage Cited in: Acta Biochim Biophys Sin (Shanghai). 2018 Feb 1;50(2):144-155.

    Autophagy is involved in Rg1-inhibited apoptosis in macrophages Macrophages with serum deprivation are treated with 50 μM Rg1 for 48 h in the absence or presence of 3-MA (5 mM). The protein expression level of cleaved caspase-3 determined by western blot analysis in Raw264.7 macrophages.

    3-Methyladenine purchased from MCE. Usage Cited in: Acta Biochim Biophys Sin (Shanghai). 2018 Feb 1;50(2):144-155.

    Raw264.7 macrophages with serum deprivation are treated with 50 μM Rg1 for 48 h in absence or presence of compound C (10 mM) or AICAR (250 μM). Western blots of the protein expressions of Atg5, Beclin1, LC3, and p62/SQSMT1.

    3-Methyladenine purchased from MCE. Usage Cited in: Acta Biochim Biophys Sin (Shanghai). 2018 Feb 1;50(2):144-155.

    Raw264.7 macrophages treated without or with Rapamycin (1 μΜ) or Chloroquine (20 μΜ) for 48 h. Western blot shows the protein expression levels of Atg5, Beclin1, LC3, and p62/SQSMT1.

    3-Methyladenine purchased from MCE. Usage Cited in: Oncotarget. 2017 Nov 22;8(65):109135-109150.

    MCF-7 cells are pretreated with the indicated chemical inhibitors for 30min, followed by 15 min treatment with RA (20 μM) + EPA (80 μM).Cell extracts are prepared and subjected to western blotting analysis.

    3-Methyladenine purchased from MCE. Usage Cited in: Oncotarget. 2017 Nov 22;8(65):109135-109150.

    Cell morphology of MCF-7 treated with RA (20 μM)+ω-3 PUFAs (80 μM) with or without 3-MA (5 mM) for 24h.

    3-Methyladenine purchased from MCE. Usage Cited in: Oncotarget. 2017 Nov 22;8(65):109135-109150.

    Cells are treated with RA (20 μM) plus ω-3 PUFAs (80 μM) with or without CQ (5 μM) for 24 h. Cell extracts are prepared and subjected to western blotting analysis.

    3-Methyladenine purchased from MCE. Usage Cited in: Chem Biol Interact. 2018 Mar 1;283:59-74.

    The expression levels of apoptosis-associated proteins after treatment with morusin or tumor necrosis factor-alpha (TNF-α)/Cycloheximide (CHX). A2780 cells are incubated with the indicated concentrations of morusin or TNF-α (50 ng/mL)/CHX (50 μM) for 24 h. GAPDH is used as a loading control in western blot analysis.

    3-Methyladenine purchased from MCE. Usage Cited in: Chem Biol Interact. 2018 Mar 1;283:59-74.

    The expression levels of apoptosis-associated proteins after treatment with morusin or tumor necrosis factor-alpha (TNF-α)/Cycloheximide (CHX). SKOV-3 cells are incubated with the indicated concentrations of morusin or TNF-α (50 ng/mL)/CHX (50 μM) for 24 h. GAPDH is used as a loading control in western blot analysis.

    3-Methyladenine purchased from MCE. Usage Cited in: Cell Physiol Biochem. 2018;48(6):2318-2336.

    Cells are pretreated with 5 mM 3-MA or 2 μM for 1 h and then exposed to 40 μM EPA and 10 μM Rp for 48 h. Cell extracts are prepared and subjected to western blot analysis.

    3-Methyladenine purchased from MCE. Usage Cited in: Cell Physiol Biochem. 2018;48(6):2318-2336.

    Cells are treated with Rapamycin (Rp; 10 μM)+Eicosapentaenoic acid (EPA; 40 μM) with or without Chloroquine (CQ; 5 μM) for 48 h. Cell extracts are prepared and subjected to western blot analysis.

    3-Methyladenine purchased from MCE. Usage Cited in: Biochem Biophys Res Commun. 2016 Dec 2;481(1-2):90-96.

    The hepatic stellate cells (HSCs) activation is down-regulated along with autophagy following DMKG or 3MA treatment in vitro. Western blot analysis of LC3B, Beclin-1, α-SMA, and collagen-I expression from lysates of HSC-T6 incubated with 4mM DMKG or 10 mM 3MA for 12 h.

    3-Methyladenine purchased from MCE. Usage Cited in: Biochem Biophys Res Commun. 2016 Dec 2;481(1-2):90-96.

    DMKG inhibits autophagy of HSCs via acetyl-coenzyme A and EP300. Western blot analysis of LC3B, Beclin-1, a-SMA, and collagen-I expression from lysates of HSC-T6 incubated with control media, DMKG (4 mM), or Lipoic acid (5 mM) in the presence or absence of C646 (10 mM) for 12 h.

    3-Methyladenine purchased from MCE. Usage Cited in: J Dermatol Sci. 2018 Jan;89(1):11-18.

    Inhibitory effects of LED 585 nm on melanogenesis is reversed by 3-MA.

    3-Methyladenine purchased from MCE. Usage Cited in: J Dermatol Sci. 2018 Jan;89(1):11-18.

    Inhibitory effects of LED 585 nm on melanogenesis is reversed by 3-MA. Autophagy is decreased in 3-MA-treated group, which is reflected by the decrease in the ratio of LC3 II/LC3 I and increase in p62 protein

    3-Methyladenine purchased from MCE. Usage Cited in: Phytother Res. 2018 Jul;32(7):1320-1331.

    Panc-28 cells are treated with GGA (20 μM) for 48 hr and the expression of HSPA8 is determined using western blot.

    3-Methyladenine purchased from MCE. Usage Cited in: Phytother Res. 2018 Jul;32(7):1320-1331.

    Panc-28 cells are treated with VER-155008 (20 μM) for 48 hr, and the expression of HSPA8 is determined using western blot

    3-Methyladenine purchased from MCE. Usage Cited in: Int J Mol Med. 2018 Jun;41(6):3221-3230.

    Western blot analysis is used to measure LC3, p62 and Beclin‑1 expression following the treatment of H9C2 cells with si‑miRNA‑30e and 3‑MA.

    3-Methyladenine purchased from MCE. Usage Cited in: Biochem Pharmacol. 2018 Mar 15;152:45-59.

    Effects of NF-κB inhibitor (10 μM MLN120B) is measured on p62 protein in LPS-induced RAW264.7 cells.

    3-Methyladenine purchased from MCE. Usage Cited in: Biochem Pharmacol. 2018 Mar 15;152:45-59.

    PFK2 expression is significantly increased by LPS but is attenuated by Canagliflozin (CAN).

    3-Methyladenine purchased from MCE. Usage Cited in: Biochem Pharmacol. 2018 Mar 15;152:45-59.

    Effects of Dapagliflozin and Empagliflozin on p62 expression in LPS-untreated and LPS-treated RAW264.7 cells.

    3-Methyladenine purchased from MCE. Usage Cited in: Biomed Res. 2018;39(2):87-94.

    Western blots are performed to determine LC3B and p62 levels in the livers to identify the autophagy status. Accumulation of LC3B II and p62 proteins in the livers of chloroform-treated mice is reduced by 3-MA treatment.

    3-Methyladenine purchased from MCE. Usage Cited in: Biomed Res. 2018;39(2):87-94.

    Accumulation of LC3B II and p62 proteins in the livers of chloroform-treated mice is reduced by 3-MA treatment.

    3-Methyladenine purchased from MCE. Usage Cited in: PLoS One. 2018 Aug 6;13(8):e0201920.

    The pretreatment of Z-VAD-FMK significantly reduces the levels of cleaved caspase-3 and cleaved PARP. The pretreatment of 3-MA blocks the conversion of LC3-I to LC3-II.

    3-Methyladenine purchased from MCE. Usage Cited in: J Immunol Res. 7 August 2018.

    Western blot assays show that 3-MA abrogates the Alda-1 induced expression increases in LC3BⅡ and Bcl2 and decreases in P62 and Bax.

    3-Methyladenine purchased from MCE. Usage Cited in: J Immunol Res. 7 August 2018.

    Western blot analysis of p-AMPK, AMPK, Bcl2 and Bax, LC3B and P62 expression in liver tissues.
    • Biological Activity

    • Protocol

    • Technical Information

    • Purity & Documentation

    • References

    Description

    3-Methyladenine is a potent inhibitor of autophagy and PI3K, blocking autophagy through its effect on PI3K, with IC50s of 25 μM and 60 μM for Vps34 and PI3Kγ, respectively.

    IC50 & Target[1]

    PtdIns3Kγ

    60 μM (IC50, Cell Assay)

    Vps34

    25 μM (IC50, Cell Assay)

    Autophagy

     

    Mitophagy

     

    Human Endogenous Metabolite

     

    In Vitro

    3-Methyladenine shows slight preference to binds to Vps34 in vitro, with an IC50 of 25 µM for Vps34 as compared with 60 µM for PtdIns3Kγ, and 3-Methyladenine (10 mM) can inhibit all PtdIns3Ks[1]. 3-Methyladenine (3-MA, 5 mM) suppresses autophagy in HeLa cells under both glucose-free conditions and normal conditions. 3-Methyladenine (2.5, 5 or 10 mM) induces caspase-dependent cell death in HeLa cells, and the death occurs independently of the inhibition of autophagy. Moreover, 3-Methyladenine (3-MA, 1 mM) significantly shortens the duration of nocodazole-induced-prometaphase arrest[2].

    In Vivo

    3-Methyladenine (1.5 mg/100 g, i.p.) treatment alleviates sodium taurocholate-induced severe acute pancreatitis (SAP) in rats at both 12 and 24 h. 3-Methyladenine inhibits autophagy of pancreatic acinar cells in sodium taurocholate-tnduced SAP. 3-Methyladenine also shows inhibitory effects on PI3K/Akt signaling pathway and NF-κB signaling pathway in sodium taurocholate-tnduced SAP[3].

    Solvent & Solubility
    In Vitro: 

    H2O : 8.82 mg/mL (59.14 mM; Need ultrasonic)

    DMSO : ≥ 1.5 mg/mL (10.06 mM)

    *"≥" means soluble, but saturation unknown.

    Preparing
    Stock Solutions
    Concentration Solvent Mass 1 mg 5 mg 10 mg
    1 mM 6.7047 mL 33.5233 mL 67.0466 mL
    5 mM 1.3409 mL 6.7047 mL 13.4093 mL
    10 mM 0.6705 mL 3.3523 mL 6.7047 mL
    *Please refer to the solubility information to select the appropriate solvent.
    In Vivo:
    • 1.

      3-Methyladenine is prepared in PBS[4].

    References
    Cell Assay
    [2]

    Cell viability is determined by a trypan blue exclusion assay. Cells are cultured in the medium with 3-Methyladenine. Both adherent and floating cells are collected and suspended in phosphate buffered saline (PBS, pH 7.4) at a final density of 1-2×106/mL. An equal volume of 0.4% trypan blue solution (w/v, in PBS) is added to the cell suspension and mixed thoroughly. After incubation at room temperature for 3 min, cell counting is performed using a hemacytometer.

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

    Animal Administration
    [3]

    All rats are fasted for 12 h with free access to water prior to operation. After anesthesia by intraperitoneal (i.p.) injection of 2% sodium pentobarbital (0.25 mL/100 g), they are laid and fixed on the table, routinely shaven, disinfected, and draped. The rat SAP model is induced by 0.1 mL/min speed uniformly retrograde infusion of a freshly prepared 3.5% sodium taurocholate solution (0.1 mL/100 g) into the biliopancreatic duct after laparotomy. Equivalent volume of normal saline solution is substituted for 3.5% sodium taurocholate solution in the sham-operation (SO) control group. The incision is closed with a continuous 3-0-silk suture, and 2 mL/100 g of saline is injected into the back subcutaneously to compensate for the fluid loss. 180 rats are randomly divided into four groups: (1) Acanthopanax treatment group (Aca group, n = 45) where the rats are injected with 0.2% Acanthopanax injection at a dose of 3.5 mg/100 g 3 h after successful modeling via the vena caudalis once, knowing that this dosage is effective; (2) 3-Methyladenine treatment group (3-methyladenine group, n = 45) where the rats are injected with 100 nmol/μL 3-methyladenine solution at a dose of 1.5 mg/100 g 3 h after successful modeling via the intraperitoneal route once, knowing that this dosage is effective; (3) SAP model group (SAP group, n = 45) where these rats receive an equivalent volume of the normal saline instead of Acanthopanax injection 3 h after successful modeling via the vena caudalis once; (4) SO group (control, n = 45) where these rats receive an equivalent volume of the normal saline instead of Acanthopanax injection 3 h after successful sham-operation via the vena caudalis once. The 45 animals in each of the four groups are equally randomized into 3, 12, and 24 h subgroups for postoperative observations[3].

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

    References
    Molecular Weight

    149.15

    Formula

    C₆H₇N₅

    CAS No.

    5142-23-4

    SMILES

    NC1=C2N=CN=C2N(C)C=N1

    Storage
    Powder -20°C 3 years
      4°C 2 years
    In solvent -80°C 6 months
      -20°C 1 month
    Shipping

    Room temperature in continental US; may vary elsewhere

    Purity: 99.84%

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    Product Name:
    3-Methyladenine
    Cat. No.:
    HY-19312
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    3-Methyladenine

    Cat. No.: HY-19312