A-769662

Catalog No.S2697

A-769662 Chemical Structure

Molecular Weight(MW): 360.39

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

Size Price Stock Quantity  
In DMSO USD 210 In stock
USD 147 In stock
USD 270 In stock
USD 470 In stock
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Cited by 15 Publications

4 Customer Reviews

  • Cells were treated with CP, DOXO, SRT1720 (100 nM), EX527 (100 nM), A769662 (10 μM) and Compound C (1 μM) under normoxic or hypoxic conditions for 48 hours, and then their viabilities were measured by MTT.

    Cancer Res 2014 74(1), 298-308. A-769662 purchased from Selleck.

  • Concentration-dependent effect of synthetic AMPK stimulator, A-769662, applied for 10 min on phosphorylated α subunit of AMPK. AMPK inhibitor (compound C, CC) was added to some samples at 10 uM. **p < 0.01, ***p < 0.001 vs. sample without A-769662 by ANOVA and Tukey post hoc test.

    Pharmacol Res 2014 81, 34-43. A-769662 purchased from Selleck.

  • Eight- to 10-week-old Ldlr-/- mice fed a standard laboratory chow were fasted overnight, fed at 07:00 h for 2 h, and refasted at 09:00 h. Intraperitoneal injection of vehicle, GW1516 (3 mg/kg), or A-769662 (30 mg/kg) (n = 6/group) occurred at the beginning of the refasting period at 09:00 h. Immunoblots of AMPK and ACC in freeze-clamped liver lysates 90 min postinjection. Representative immunoblots with quantitations are shown. Asterisk (*) indicates significant difference between vehicle and treatment; Student's paired t-test (P < 0.05).

    J Lipid Res 2014 55(7), 1254-1266. A-769662 purchased from Selleck.

  • Cultured PANC-1 pancreatic cancer cells were treated with vehicle (DMSO, 1%), 10 uM of belinostat (BS, for 2 h, 4 h and 6 h) or A-769662 (10 uM, 2 h), phospho- and total AMPK and ACC were detected by western blot, tubulin (loading control) was also tested. AMPK and ACC phosphorylation was quantified as described.

    Biochem Biophys Res Commun 2013 437(1), 1-6. A-769662 purchased from Selleck.

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 M2nzPWZ2dmO2aX;uJIF{e2G7 M1;jcFEhdU1? Mlq5SG1UVw>? NFG2ZotqdmirYnn0d{Bn[XS2eTDhZ4llKHO7boTo[ZNqeyC5aYToJGlEPTBib3[gN{43KM7:TR?= M{fVfVE3PzV|NUe2
rat hepatocytes M{e2WWZ2dmO2aX;uJIF{e2G7 MXqxJI1O NEfVUplFVVOR MXHpcohq[mm2czDmZZR1gSCjY3nkJJN6dnSqZYPpd{B4cXSqIFnDOVAhd2ZiMz62JO69VQ>? M4PVTFE3PzV|NUe2
HEK293 M33oPGtqdmG|ZTDhd5NigQ>? NE\KNWMzODBizszN MVPEUXNQ MWPhZ5RqfmG2ZYOg[Y5ld2enbn;1d{BCVVCN M4WxdFE4PzJ6MkSx
CCL13 M3Hne2tqdmG|ZTDhd5NigQ>? NF;TVlIzODBizszN MVjEUXNQ NIrNPGVi[3SrdnH0[ZMh\W6mb3flco92eyCDTWDL NIPVVJoyPzd{OEK0NS=>
MEFs MnfuSpVv[3Srb36gZZN{[Xl? MX6zNFAh|ryP MX\EUXNQ M3nmOIlvcGmkaYTzJJBzd3SnYYPvcYFtKG[3bnP0bY9vKGK7IHHuJGFOWEtvaX7k[ZBmdmSnboSgcYVkcGGwaYPt MlnkNVg2QTN3OES=
epididymal clear cells MV3GeY5kfGmxbjDhd5NigQ>? M{HB[FIxOCEQvF2= NF\uPGxFVVOR M1nlOYlvcGmkaYTzJJRp\SCySD3t[YRq[XSnZDDWMWFVWGG|ZTDhZ4N2dXWuYYTpc44h[XRidHjlJIFxcWOjbDDt[Y1jemGwZR?= NV;zc3JNOTl{MUG5NVg>
3T3-L1 MlHHSpVv[3Srb36gZZN{[Xl? MlH5NU4zKG2P MV;EUXNQ MUDpcohq[mm2czCzWFMuVDFiQXTpdI9o\W6nc3nz NXzreIVIOTl2OEOzNFQ>
3T3-L1 MmPwSpVv[3Srb36gZZN{[Xl? NEj0T2wyNjJibV2= M1\IemROW09? NIrjZXJqdmirYnn0d{B1cGViRYjwdoV{e2mxbjDv[kBC\Gmyb3flcoV{cXOUZXzheIVlKFS{YX7zZ5JqeHSrb36gSoFkfG:{czDhcoQhVWG{a3Xydy=> MYmxPVQ5OzNyNB?=
3T3-L1 MV3GeY5kfGmxbjDhd5NigQ>? NF\OcGcyNjJibV2= M{T1SGROW09? MnHBbY5pcWKrdIOgUYl1d3SrYzDDcI9v[WxiRYjwZY5{cW:w M1fhPVE6PDh|M{C0
3T3-L1 M2fxUoN6fG:2b4jpZ4l1gSCjc4PhfS=> NVzmT|ZWOS5{IH3N MlW1SG1UVw>? NVrOeIxF\GWlcnXhd4V{KEOnbHygWoli[mmuaYT5 NEXUepkyQTR6M{OwOC=>
3T3-L1 MlPkT4lv[XOnIHHzd4F6 Mnq1NU4zKG2P MVzEUXNQ M4XPb4FkfGm4YYTld{BCVVCN MmfONVk1QDN|MES=
L6 skeletal muscle cells Mk\GSpVv[3Srb36gZZN{[Xl? MkLhNlUxKM7:TR?= NFvZbY9FVVOR M4nGVYFkfGm4YYTld{BCVVCNIIPp[45idGmwZzDwZZRpf2G7cx?= NFXYfosyQTh{OEizOi=>
L6 skeletal muscle cells NFLZUphHfW6ldHnvckBie3OjeR?= NX2zflYxOjVyIN88US=> NF6zUWNFVVOR MmThbY5pcWKrdIOgeIhmKE6jKz3LL{1CXFCjc3WgeJJidnOyb4L0JIFkfGm4aYT5JIFv\CClZXzsJJN2em[jY3WgZYJ2dmSjbnPl NWDYZZJiOTl6Mki4N|Y>
MDA-MB231 NGHpS5JCeG:ydH;zbZMh[XO|YYm= Mn6yOFAxKM7:TR?= NFvYd3dFVVOR Mlz5d4Vve2m2aYrld{BpfW2jbjDidoVie3RiY3HuZ4VzKGOnbHygcIlv\XNidH:gWHJCUUxvaX7keYNm\CCjcH;weI9{cXN? M3vqeFE6QDl4NE[5
BT474 MojKRZBweHSxc3nzJIF{e2G7 NEjEd4c1ODBizszN MV3EUXNQ M2TtVpNmdnOrdHn6[ZMhcHWvYX6gZpJm[XO2IHPhcoNmeiClZXzsJIxqdmW|IITvJHRTSUmOLXnu[JVk\WRiYYDvdJRwe2m| NXTt[HdLOTl6OU[0Olk>
MCF7 MoTSRZBweHSxc3nzJIF{e2G7 MYO0NFAh|ryP MmLVSG1UVw>? MV3z[Y5{cXSrenXzJIh2dWGwIHLy[YF{fCClYX7j[ZIh[2WubDDsbY5meyC2bzDUVmFKVC2rbnT1Z4VlKGGyb4D0c5Nqew>? MVKxPVg6PjR4OR?=
Mesenchymal stem cells NYLuPHAxU2mwYYPlJIF{e2G7 MXKxNEDDvU1? MYDEUXNQ MYnpcoR2[2W|IHGgdo9jfXO2IHHu[EB{fXO2YXnu[YQhSU2SSzDhZ5RqfmG2aX;u NH;MTGUzPDFyNEi3PS=>
Mesenchymal stem cells MkW3Z5l1d3SxeHnjbZR6KGG|c3H5 MkO1NVAxKML3TR?= NX\hTYMyTE2VTx?= NVzqXWFN\GWlcnXhd4V{KHSqZTDNV2MheHKxbHnm[ZJifGmxbh?= M4r3R|I1OTB2OEe5
MG-63 NHXUPJNkgXSxdH;4bYNqfHliYYPzZZk> M4L3SVExKML3TR?= MoK0SG1UVw>? NXvWUoJwcW6qaXLpeJMhUDKRMj3JcoR2[2WmIF;zeIVw[myjc4SgR4VtdCCGZXH0bC=> MkTwNlQ6PjB|NkK=
MC3T3-E1 NE[zZlFkgXSxdH;4bYNqfHliYYPzZZk> NGHzNFMyOCEEtV2= NIe1UYdFVVOR MoTJbY5pcWKrdIOgTFJQOi2LbnT1Z4VlKE:|dHXvZoxie3RiQ3XscEBF\WG2aB?= NFzGNVYzPDl4MEO2Ni=>
MG-63 NIjPdW9CeG:ydH;zbZMh[XO|YYm= NVTqUY1oOTBiwsXN MnnNSG1UVw>? M{DlVpN2eHC{ZYPz[ZMhUDKRMj3JcoR2[2WmIF;zeIVw[myjc4SgR4VtdCCDcH;weI9{cXN? MWqyOFk3ODN4Mh?=
MC3T3-E1 NHTUfZNCeG:ydH;zbZMh[XO|YYm= MnXpNVAhyrWP MkeySG1UVw>? NYrEfmdGe3WycILld5NmeyCKMl:yMWlv\HWlZXSgU5N1\W:kbHHzeEBE\WyuIFHwc5B1d3Orcx?= MnnENlQ6PjB|NkK=
MG-63 M1fZOmZ2dmO2aX;uJIF{e2G7 NFn3XFUyOCEEtV2= MWLEUXNQ MWXhcIxmfmmjdHXzJHJQWyCjY3P1cZVt[XSrb36gZY5lKEGWUDDk[ZBt\XSrb36gZ4F2e2WmIHL5JGgzVzJ? MWOyOFk3ODN4Mh?=
MC3T3-E1 MUfGeY5kfGmxbjDhd5NigQ>? M3zWNlExKML3TR?= M1nwdmROW09? M334XoFtdGW4aXH0[ZMhWk:VIHHjZ5VufWyjdHnvckBidmRiQWTQJIRmeGyndHnvckBk[XW|ZXSgZpkhUDKRMh?= M1PYNFI1QTZyM{[y
MG-63 M3;FSGZ2dmO2aX;uJIF{e2G7 Mn7vNVAhyrWP NV\tTHNFTE2VTx?= M{m2dYZi[2muaYTheIV{KEh{T{KtbY5lfWOnZDDheZRweGijZ4mgZYN1cX[jdHnvci=> NHTt[2IzPDl4MEO2Ni=>
MC3T3-E1 MX3GeY5kfGmxbjDhd5NigQ>? NIfnXZoyOCEEtV2= NHTGcY5FVVOR MkPo[oFkcWyrdHH0[ZMhUDKRMj3pcoR2[2WmIHH1eI9xcGGpeTDhZ5RqfmG2aX;u Mom5NlQ6PjB|NkK=
PC3 Mn71T4lv[XOnIHHzd4F6 NHyxXo4yODBiwsXN M3PnR2ROW09? MmP2eZBz\We3bHH0[ZMhfGinIHzleoVteyCxZjDBUXBMKGGwZDDBR2MheGixc4Doc5J6dGG2aX;u NGrTdpIzPTV7NEC0Ny=>
PC3M NVfOe4J3U2mwYYPlJIF{e2G7 MVqxNFAhyrWP M1fQPWROW09? MlfweZBz\We3bHH0[ZMhfGinIHzleoVteyCxZjDBUXBMKGGwZDDBR2MheGixc4Doc5J6dGG2aX;u MlrDNlU2QTRyNEO=
PC3 NGXCcFBHfW6ldHnvckBie3OjeR?= M1zhTVExOCEEtV2= Mo\NSG1UVw>? NXXH[ll6cW6mdXPld{BRUTONL33UU3IheGG2aIfhfZM> M2LI[lI2PTl2MESz
PC3M MkD4SpVv[3Srb36gZZN{[Xl? M2fGZ|ExOCEEtV2= MYjEUXNQ MUjpcoR2[2W|IGDJN2swdVSRUjDwZZRpf2G7cx?= NUPz[m5SOjV3OUSwOFM>
PC3 M{HKOWdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 MUKxNFAhyrWP Ml;YSG1UVw>? MmHUd5VxeHKnc4Pld{Bxem:uaX\ldoF1cW:w NHfte|gzPTV7NEC0Ny=>
PC3M MnHJS5Jwf3SqIHnubIljcXSxcomgZZN{[Xl? MXixNFAhyrWP M3TJ[WROW09? MX3zeZBxemW|c3XzJJBzd2yrZnXyZZRqd25? MWCyOVU6PDB2Mx?=
MC3T3-E1 Ml3UT4lv[XOnIHHzd4F6 M{HCTVExKM7:TR?= M{i1VWROW09? MUfpcoR2[2W|IIPp[45q\mmlYX70JGFOWEtiYXP0bZZifGmxbh?= NYHKWVNYOjZ6OUG4OlY>
MC3T3-E1 M17wXmdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 M1jOUVExKM7:TR?= MULEUXNQ MWHpcohq[mm2czDE[ZgucW6mdXPl[EBwe3Snb3LsZZN1KGOnbHyg[IVifGh? M1fNOlI3QDlzOE[2
MC3T3-E1 NXHsfJhWTnWwY4Tpc44h[XO|YYm= Ml;ONVAh|ryP NWnJZVA3TE2VTx?= NVe5WGNmcW6qaXLpeJMhTGW6LXnu[JVk\WRib4jp[IF1cX[nIIP0doV{ew>? MX[yOlg6OTh4Nh?=

... Click to View More Cell Line Experimental Data

Assay
Methods Test Index PMID
Western blot
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.

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䲧疝Ỵ疞㧀疜膉痘 瘿뾠ՂᾰƌՂĀ 㺣痖帉痖Ѐ瑖堘𢡄빢᎒ՂĀ鑸᎒彿堙奋堙巫堙᎒ﻺ᎒彿堙ﻮ᎒塚堙ﻺ᎒ꍈ堞빢᎒學堙漸堞圔堙빢᎒圞堙圭堙𢡄玚Wᾰƌ ᾰƌ戤瘯Ɖ뙠ෆ䐺痖暼瘿뙠ෆᾰƌ 뙠ෆÐ㺣痖뙠ෆ€𢡄뙤ෆ€䀷痗뙤ෆ౴뙤ෆ㵶痗뙤ෆ뺖᎒泌

Cleaved caspase-8 / Cleaved caspase-9 / Cleaved caspase-3 ; 

PubMed: 31193061     


(C) Caspase activation was determined by western blotting. Flavopiridol treatment (300 nM) induced the expressions of cleaved caspase-8, -9 and -3 in a time-dependent manner. 

p-RNAPII / p-eIF4E / Mnk1 ; 

PubMed: 24572052     


After 24 h drug exposure, CDKI-73 or flavopiridol also abolished p-eIF4E(S209) at 0.25 μM, indicating cellular inhibition of Mnk kinase activity. The same treatment with CDKI-73 or flavopiridol caused a loss in Mnk1 protein expression. CGP57380-treated cells abrogated p-RNAPIIS2 as well as p-eIF4E(S209) with a minimal effect on Mnk1 protein level.

p-ERK / ERK / p-p38 / p-4EBP1 / 4EBP1 / p-S6 ; 

PubMed: 24572052     


0.25 μM CDKI-73 or flavopiridol caused little changes in the phosphorylation of Erk and p38 MAPK; however, inhibited the 4E-BP1 phosphorylation [p-4E-BP1(Thr70)] by 24 h. CGP57380 had a minimal effect on these proteins.

CDK2 / CDK4 / Cyclin A / p21 / p27 / Rb ; 

PubMed: 24572052     


Effect of flavopiridol on cell cycle-related protein expression in uterine leiomyoma cells. Twenty-four hours after treatment, cell extracts were prepared and subjected to immunoblotting analysis. β-Actin was used as an internal loading control.

22456226 26431377 31193061 24572052
Growth inhibition assay
Cell viability; 

PubMed: 31193061     


The antiproliferative effect of flavopiridol on CCA cell lines was determined using an MTT assay. KKU-055, KKU-100, KKU-213 and KKU-214 cells were treated with 50, 100, 200 or 300 nM of flavopiridol at 24, 48 or 72 h. The percentage of cell number in vehicle control was taken as 100%. Data are mean ± SD of three independent experiments. *P < 0.05 in all CCA cell lines, significantly different for each time point compared with vehicle control.

31193061
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]

+ Expand

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]

+ Expand
  • 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]

+ Expand
  • Animal Models: Sprague Dawley rats
  • Formulation: A-769662 is dissolved in DMSO.
  • 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

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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.

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID