A-769662

For research use only.

Catalog No.S2697

51 publications

A-769662 Chemical Structure

CAS No. 844499-71-4

A-769662 is a potent, reversible AMPK activator with EC50 of 0.8 μM in cell-free assays, little effect on GPPase/FBPase activity.

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Selleck's A-769662 has been cited by 51 publications

Purity & Quality Control

Choose Selective AMPK Inhibitors

Biological Activity

Description A-769662 is a potent, reversible AMPK activator with EC50 of 0.8 μM in cell-free assays, little effect on GPPase/FBPase activity.
Targets
AMPK [1]
(Cell-free assay)
Fatty acid synthesis [1]
(in primary rat hepatocytes)
0.8 μM(EC50) 3.2 μM
In vitro

A-769662 stimulates partially purified rat liver AMPK with EC50 with 0.8 μM. A-769662 activates AMPK purified from multiple tissues and species in a dose-responsive manner with modest variations in observed EC50s. EC50s determined for A-769662 using partially purified AMPK extracts from rat heart, rat muscle, or human embryonic kidney cells (HEKs) are 2.2 mM, 1.9 mM, or 1.1 mM, respectively. A 4 hours treatment of primary rat hepatocytes with A-769662 dose-dependently increases ACC phosphorylation, which correlated inhibition of fatty acid synthesis with IC50 of 3.2 μM. A-769662 also inhibits fatty acid sythesis in mouse hepatocytes with IC50 with 3.6 μM [1] A-769662 activates AMPK both allosterically and by inhibiting dephosphorylation of AMPK on Thr-172, similar to the effects of AMP. [2] A-769662 inhibits proteasomal function by an AMPK-independent mechanism. A-769662 affects the in vitro activity of purified 26S proteasomes but not the in vitro activity of purified 20S proteasomes. A-769662 has toxic effects on MEF cells. [3] A recent research shows A-769662 inhibited cell proliferation and DNA synthesis. [4]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
mouse hepatocytes M2TqN2Z2dmO2aX;uJIF{e2G7 NXfNSm1LOSCvTR?= NETLNFVFVVOR M3jQbIlvcGmkaYTzJIZifHS7IHHjbYQhe3mwdHjld4l{KHerdHigTWM2OCCxZjCzMlYh|ryP MVG8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8yPjd3M{W3Okc,OTZ5NUO1O|Y9N2F-
rat hepatocytes MmjXSpVv[3Srb36gZZN{[Xl? MVOxJI1O NWHXWHB2TE2VTx?= NFK0R4ZqdmirYnn0d{Bn[XS2eTDhZ4llKHO7boTo[ZNqeyC5aYToJGlEPTBib3[gN{43KM7:TR?= MlnTQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOTZ5NUO1O|YoRjF4N{WzOVc3RC:jPh?=
HEK293 MlTZT4lv[XOnIHHzd4F6 MWCyNFAh|ryP M3PDcWROW09? NYrTcoJZ[WO2aY\heIV{KGWwZH;n[Y5wfXNiQV3QTy=> NUTFZ4V6RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMUe3NlgzPDFpPkG3O|I5OjRzPD;hQi=>
CCL13 M1LzTGtqdmG|ZTDhd5NigQ>? NUThNY5yOjByIN88US=> MnvUSG1UVw>? M3\DS4FkfGm4YYTld{BmdmSxZ3Xuc5V{KEGPUFu= MWO8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8yPzd{OEK0NUc,OTd5MkiyOFE9N2F-
MEFs MWnGeY5kfGmxbjDhd5NigQ>? NUHpPWJ4OzByIN88US=> M4rzeGROW09? NF[2VXVqdmirYnn0d{Bxem:2ZXHzc41idCCodX7jeIlwdiCkeTDhckBCVVCNLXnu[IVx\W6mZX70JI1m[2ijbnnzcS=> M4fWWFxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzF6NUmzOVg1Lz5zOEW5N|U5PDxxYU6=
epididymal clear cells MX;GeY5kfGmxbjDhd5NigQ>? M4rPZ|IxOCEQvF2= MmW1SG1UVw>? NWX5SVJRcW6qaXLpeJMhfGinIIDIMY1m\GmjdHXkJHYuSVSSYYPlJIFk[3WvdXzheIlwdiCjdDD0bIUh[XCrY3HsJI1mdWK{YX7l Mn34QIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOTl{MUG5NVgoRjF7MkGxPVE5RC:jPh?=
3T3-L1 M{LmcmZ2dmO2aX;uJIF{e2G7 M3LTRVEvOiCvTR?= MoWySG1UVw>? M1nTV4lvcGmkaYTzJFNVOy2OMTDB[Ilxd2enbnXzbZM> NWrEd2VlRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMUm0PFM{ODRpPkG5OFg{OzB2PD;hQi=>
3T3-L1 MVXGeY5kfGmxbjDhd5NigQ>? M4DYOlEvOiCvTR?= MXTEUXNQ NFnZVZFqdmirYnn0d{B1cGViRYjwdoV{e2mxbjDv[kBC\Gmyb3flcoV{cXOUZXzheIVlKFS{YX7zZ5JqeHSrb36gSoFkfG:{czDhcoQhVWG{a3Xydy=> NVfrXpprRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMUm0PFM{ODRpPkG5OFg{OzB2PD;hQi=>
3T3-L1 M{HW[mZ2dmO2aX;uJIF{e2G7 MWexMlIhdU1? M3LSW2ROW09? NWr4Vmt1cW6qaXLpeJMhVWm2b4TpZ{BEdG:wYXygSZhx[W6|aX;u MXK8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8yQTR6M{OwOEc,OTl2OEOzNFQ9N2F-
3T3-L1 NVPFeFZp[3m2b4TvfIlkcXS7IHHzd4F6 NFu2NI0yNjJibV2= M1LZfWROW09? M1PEeYRm[3KnYYPld{BE\WyuIG\pZYJqdGm2eR?= MVO8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8yQTR6M{OwOEc,OTl2OEOzNFQ9N2F-
3T3-L1 MlnMT4lv[XOnIHHzd4F6 M4r2UFEvOiCvTR?= NYjFbG5GTE2VTx?= MkjpZYN1cX[jdHXzJGFOWEt? MXi8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8yQTR6M{OwOEc,OTl2OEOzNFQ9N2F-
L6 skeletal muscle cells MWHGeY5kfGmxbjDhd5NigQ>? NVTNflYzOjVyIN88US=> MYrEUXNQ MVLhZ5RqfmG2ZYOgRW1RUyC|aXfuZYxqdmdicHH0bJdigXN? MkiwQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOTl6Mki4N|YoRjF7OEK4PFM3RC:jPh?=
L6 skeletal muscle cells MYTGeY5kfGmxbjDhd5NigQ>? MlPoNlUxKM7:TR?= M{TmOWROW09? NVTXbHJxcW6qaXLpeJMhfGinIF7hL{1MMy2DVGDhd4UhfHKjboPwc5J1KGGldHn2bZR6KGGwZDDj[YxtKHO3cn\hZ4Uh[WK3bnThcoNm MkTQQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOTl6Mki4N|YoRjF7OEK4PFM3RC:jPh?=
MDA-MB231 M3zaU2Fxd3C2b4Ppd{Bie3OjeR?= MWC0NFAh|ryP NVnXWpNiTE2VTx?= MlHXd4Vve2m2aYrld{BpfW2jbjDidoVie3RiY3HuZ4VzKGOnbHygcIlv\XNidH:gWHJCUUxvaX7keYNm\CCjcH;weI9{cXN? MlvHQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOTl6OU[0OlkoRjF7OEm2OFY6RC:jPh?=
BT474 Mn\PRZBweHSxc3nzJIF{e2G7 NYj6OoU4PDByIN88US=> MYXEUXNQ NU\Zdopze2Wwc3n0bZpmeyCqdX3hckBjemWjc4SgZ4Fv[2W{IHPlcIwhdGmwZYOgeI8hXFKDSVytbY5lfWOnZDDhdI9xfG:|aYO= MnHFQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOTl6OU[0OlkoRjF7OEm2OFY6RC:jPh?=
MCF7 M3zJNWFxd3C2b4Ppd{Bie3OjeR?= NIqyNZA1ODBizszN NVzqWpFpTE2VTx?= NH61V5F{\W6|aYTpfoV{KGi3bXHuJIJz\WG|dDDjZY5k\XJiY3XscEBtcW6nczD0c{BVWkGLTD3pcoR2[2WmIHHwc5B1d3Orcx?= NELMNpU9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9zOUi5OlQ3QSd-MUm4PVY1Pjl:L3G+
Mesenchymal stem cells NV;ocFNLU2mwYYPlJIF{e2G7 MXOxNEDDvU1? Mkn4SG1UVw>? NYnUVIRvcW6mdXPld{BiKHKxYoXzeEBidmRic4XzeIFqdmWmIFHNVGsh[WO2aY\heIlwdg>? MWC8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zPDFyNEi3PUc,OjRzMES4O|k9N2F-
Mesenchymal stem cells NIH1[XdkgXSxdH;4bYNqfHliYYPzZZk> M2Dp[lExOCEEtV2= MlTTSG1UVw>? MUXk[YNz\WG|ZYOgeIhmKE2VQzDwdo9tcW[ncnH0bY9v NFLwR|U9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{NEGwOFg4QSd-MkSxNFQ5Pzl:L3G+
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MC3T3-E1 M2HnW4N6fG:2b4jpZ4l1gSCjc4PhfS=> M4fUbVExKML3TR?= MmeySG1UVw>? MnT2bY5pcWKrdIOgTFJQOi2LbnT1Z4VlKE:|dHXvZoxie3RiQ3XscEBF\WG2aB?= NUnyOoFURGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkS5OlA{PjJpPkK0PVYxOzZ{PD;hQi=>
MG-63 NUPBRWx3SXCxcITvd4l{KGG|c3H5 MnXWNVAhyrWP M2fTTWROW09? NXy0WG9we3WycILld5NmeyCKMl:yMWlv\HWlZXSgU5N1\W:kbHHzeEBE\WyuIFHwc5B1d3Orcx?= MVS8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zPDl4MEO2Nkc,OjR7NkCzOlI9N2F-
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MG-63 M1K0Z2Z2dmO2aX;uJIF{e2G7 MUmxNEDDvU1? NID0NFFFVVOR MoqzZYxt\X[rYYTld{BTV1NiYXPjeY12dGG2aX;uJIFv\CCDVGCg[IVxdGW2aX;uJINifXOnZDDifUBJOk9{ NH3leFE9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{NEm2NFM3Oid-MkS5OlA{PjJ:L3G+
MC3T3-E1 M{\OZWZ2dmO2aX;uJIF{e2G7 MWWxNEDDvU1? NEf5cnFFVVOR Mn\1ZYxt\X[rYYTld{BTV1NiYXPjeY12dGG2aX;uJIFv\CCDVGCg[IVxdGW2aX;uJINifXOnZDDifUBJOk9{ MojXQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjR7NkCzOlIoRjJ2OU[wN|YzRC:jPh?=
MG-63 Ml:5SpVv[3Srb36gZZN{[Xl? MXmxNEDDvU1? Mlr2SG1UVw>? MYnmZYNqdGm2YYTld{BJOk9{LXnu[JVk\WRiYYX0c5Bp[We7IHHjeIl3[XSrb36= NXrGPIU4RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkS5OlA{PjJpPkK0PVYxOzZ{PD;hQi=>
MC3T3-E1 MlPkSpVv[3Srb36gZZN{[Xl? M3OyWlExKML3TR?= NIPsOFBFVVOR Mlny[oFkcWyrdHH0[ZMhUDKRMj3pcoR2[2WmIHH1eI9xcGGpeTDhZ5RqfmG2aX;u MV68ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zPDl4MEO2Nkc,OjR7NkCzOlI9N2F-
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PC3M MkG5S5Jwf3SqIHnubIljcXSxcomgZZN{[Xl? MknVNVAxKML3TR?= M4XFPGROW09? M4LPT5N2eHC{ZYPz[ZMheHKxbHnm[ZJifGmxbh?= NGXNUXg9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{NUW5OFA1Oyd-MkW1PVQxPDN:L3G+
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MC3T3-E1 NF7iPXhHfW6ldHnvckBie3OjeR?= MX6xNEDPxE1? NVzlRVRzTE2VTx?= MnHHbY5pcWKrdIOgSIV5NWmwZIXj[YQhd3irZHH0bZZmKHO2cnXzdy=> M162NVxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ4OEmxPFY3Lz5{Nki5NVg3PjxxYU6=
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SNG-M M3LZbmdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 M4TwVWlvcGmkaYTpc44hd2ZiaIXtZY4hW06JLV2gZ4VtdCCpcn;3eIghcW5iYTDj[YxtKH[rYXLpcIl1gSCjc4PhfUwhUUN3MDC9JFQ5NjZ5MEGg{txONg>? NVXE[YNWRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:ve5d4NmWkaT7hZ{52cy:laHXtZoww[2:vcH;1coRgemWyb4L0Y4NiemRxQ1jFUWJNPTF{NUC0M{c,W0GQR1XSQE9iRg>?
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... Click to View More Cell Line Experimental Data

Assay
Methods Test Index PMID
Western blot
pCofilin / Cofilin / detyrosinated alpha tubulin / acetylated alpha tubulin ; 

PubMed: 26431377     


A & B. MCF-7 cells were treated with 100–400 μM of A-769662 for 4-24 hours. Protein was harvested and Western blot analysis was done to determine levels of pAMPK at threonine 172, pCofilin at serine 3, detyrosinated alpha tubulin (glu-tub), and acetylated alpha tubulin (acetyl-tub). C & D. BT-549 cells were treated with 100–400 μM A-769662 for 4–24 hours. Protein was harvested and Western blot analysis was done to determine levels of markers listed above.

p-AMPK / AMPK / p-S6K / S6K / p-ACC ; 

PubMed: 22456226     


Representative immunoblots for p-AMPK, total AMPK, p-S6K, total S6K, p-ACC and actin under control conditions (C) and in response to 50 μM A-769662 at various time points.

26431377 22456226
In vivo Short-term treatment of normal Sprague Dawley rats with A-769662 decreases liver malonyl CoA levels and the respiratory exchange ratio, VCO2/VO2, indicating an increased rate of whole-body fatty acid oxidation. Treatment of ob/ob mice with 30 mg/kg b.i.d. A-769662 decreases hepatic expression of PEPCK, G6Pase, and FAS, lowers plasma glucose by 40%, reduced body weight gain and significantly decreases both plasma and liver triglyceride levels. [1]

Protocol

Kinase Assay:

[1]

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96-well AMPK assay:

AMPK activity is measured by monitoring phosphorylation of the SAMS peptide substrate (20 mM in standard assays and 100 mM in additivity assays) following a previously described protocol (Anderson et al., 2004). To determine whether A-769662-induced AMPK activation occurs in a reversible manner, AMP or A-769662 are preincubated with rat liver AMPK for 10 minutes at 20 times standard assay concentrations prior to dilution and measurement of AMPK activity.
Cell Research:

[3]

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  • Cell lines: MEF cells
  • Concentrations: 300 μM
  • Incubation Time: 24 hours
  • Method:

    Cell viability of MEF cells treated or not with A-769662 is performed as follows: cells are harvested by trypsinization and incubated with 0.5 mg/mL RNase and 50 μg/mL propidium iodine at room temperature in the dark; cell viability is analyzed by flow cytometry using a FACScanto flow cytometer, using an excitation laser at 488 nm and a propidium iodine fluorescence detection at 600 nm. To determine the proportion of cells in each phase of the cell cycle, cells are harvested by trypsinization, collected by centrifugation, washed in PBS and fixed overnight in 80% ethanol at -20 °C. Subsequently, these fixed cells are centrifuged to remove the fixative and incubated for 20 minutes in the dark at room temperature in PBS containing 0.5 mg/mL RNase and 50 μg/mL propidium iodine. Flow cytometry analysis is performed as above. The proportion of cells in G1, S, and G2 is determined using the MODFIT program. Cell culture pictures are taken at the indicated times using a camera coupled to an inverted microscope with a 20 × objective.


    (Only for Reference)
Animal Research:

[1]

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  • Animal Models: Sprague Dawley rats
  • Dosages: 30 mg/kg
  • Administration: Administered via i.p.
    (Only for Reference)

Solubility (25°C)

In vitro DMSO 72 mg/mL (199.78 mM)
Water Insoluble
Ethanol Insoluble
In vivo Add solvents to the product individually and in order(Data is from Selleck tests instead of citations):
5% DMSO+40% PEG 300+5% Tween 80+50% ddH2O
For best results, use promptly after mixing.
5 mg/ml

* 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 360.39
Formula

C20H12N2O3S

CAS No. 844499-71-4
Storage powder
in solvent
Synonyms N/A
Smiles C1=CC=C(C(=C1)C2=CC=C(C=C2)C3=CSC4=C3C(=C(C(=O)N4)C#N)O)O

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
Dosage mg/kg Average weight of animals g Dosing volume per animal ul Number of animals
Step 2: Enter the in vivo formulation ()
% DMSO % % Tween 80 % ddH2O
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Bio Calculators

Molarity Calculator

Molarity Calculator

Calculate the mass, volume or concentration required for a solution. The Selleck molarity calculator is based on the following equation:

Mass (mg) = Concentration (mM) × Volume (mL) × Molecular Weight (g/mol)

  • Mass
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*When preparing stock solutions, please always use the batch-specific molecular weight of the product found on the via label and MSDS / COA (available on product pages).

Dilution Calculator

Dilution Calculator

Calculate the dilution required to prepare a stock solution. The Selleck dilution calculator is based on the following equation:

Concentration (start) x Volume (start) = Concentration (final) x Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2 ( Input Output )

  • C1
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    V2

* When preparing stock solutions always use the batch-specific molecular weight of the product found on the vial label and MSDS / COA (available online).

The Serial Dilution Calculator Equation

  • Serial Dilutions

  • Computed Result

  • C1=C0/X C1: LOG(C1):
    C2=C1/X C2: LOG(C2):
    C3=C2/X C3: LOG(C3):
    C4=C3/X C4: LOG(C4):
    C5=C4/X C5: LOG(C5):
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    C8=C7/X C8: LOG(C8):
Molecular Weight Calculator

Molecular Weight Calculator

Enter the chemical formula of a compound to calculate its molar mass and elemental composition:

Total Molecular Weight: g/mol

Tip: Chemical formula is case sensitive. C10H16N2O2 c10h16n2o2

Instructions to calculate molar mass (molecular weight) of a chemical compound:

To calculate molar mass of a chemical compound, please enter its chemical formula and click 'Calculate'.

Definitions of molecular mass, molecular weight, molar mass and molar weight:

Molecular mass (molecular weight) is the mass of one molecule of a substance and is expressed in the unified atomic mass units (u). (1 u is equal to 1/12 the mass of one atom of carbon-12)
Molar mass (molar weight) is the mass of one mole of a substance and is expressed in g/mol.

Molarity Calculator

Mass Concentration Volume Molecular Weight

Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

Tel: +1-832-582-8158 Ext:3

If you have any other enquiries, please leave a message.

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AMPK Signaling Pathway Map

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID