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 PI3K inhibitor. 3-Methyladenine is a widely used inhibitor of autophagy via its inhibitory effect on class III PI3K.

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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10 mM * 1 mL in Water 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: Int J Oncol. 2018 Sep;53(3):1363-1373.

    The protein expression levels of LC3 and Beclin-1 proteins in HCT-116 cells. Cells are treated with 100 μg/mL active fraction of clove (AFC) for 48 h with or without pretreatment of 2 mM 3-MA or 1 nM Baflomycin A1 (BA) for 1 h.

    3-Methyladenine purchased from MCE. Usage Cited in: J Cell Mol Med. 2018 Oct;22(10):5132-5144.

    Melatonin regulation of mitophagy activity in the diabetic myocardium, Protein expression with representative gel blots of LC3 I/II, GAPDH (loading control).

    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.

    3-Methyladenine purchased from MCE. Usage Cited in: Int J Mol Med. 2018 May;41(5):2535-2544.

    Cells are treated with AICAR and AICAR (A++, 10 μM) + Compound C (C++, 1 μM) for 24 h, after which western blot analysis is performed.

    3-Methyladenine purchased from MCE. Usage Cited in: Int J Mol Med. 2018 May;41(5):2535-2544.

    MC3T3-E1 cells are treated with AICAR (10 μM) in the presence or absence of 3-MA (5 mM) or CQ (10 μM) for 24 h, after which western blot analysis is conducted.

    3-Methyladenine purchased from MCE. Usage Cited in: J Cell Biochem. 2018 Nov 1.

    Representative Western blots show LC3 I/II and p62 protein expression in kidneys of vehicle group and 3-MA group mice at 72 hours after the Cisplatin treatment.

    3-Methyladenine purchased from MCE. Usage Cited in: Ecotox Environ Safe. 2018 Nov.

    Western blotting image protein levels of GRP78/Bip in DF-1 cells treated with different supplements (+ represent supplying, - represents not supplying a certain amount of Cr(VI), 4-PBA, Tg, NS-398, 3-MA and Rapamycin).

    3-Methyladenine purchased from MCE. Usage Cited in: Ecotox Environ Safe. 2018 Nov.

    Western blotting image protein levels of LC3-I & LC3-II in DF-1 cells treated with different supplements (+ represent supplying, - represents not supplying a certain amount of Cr(VI), 4-PBA, Tg, NS-398, 3-MA and Rapamycin).

    3-Methyladenine purchased from MCE. Usage Cited in: Ecotox Environ Safe. 2018 Nov.

    Western blotting image protein levels of p62 in DF-1 cells treated with different supplements (+ represent supplying, - represents not supplying a certain amount of Cr(VI), 4-PBA, Tg, NS-398, 3-MA and Rapamycin).
    • Biological Activity

    • Protocol

    • Technical Information

    • Purity & Documentation

    • References

    Description

    3-Methyladenine is a PI3K inhibitor. 3-Methyladenine is a widely used inhibitor of autophagy via its inhibitory effect on class III PI3K.

    IC50 & Target

    IC50: 25 μM (Vps34), 60 μM (PI3Kγ)[1]

    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