AP-III-a4 (ENOblock)

Catalog No.S7443

For research use only.

AP-III-a4 (ENOblock) is the first nonsubstrate analogue inhibitor of enolase with IC50 of 0.576 μM.

AP-III-a4 (ENOblock) Chemical Structure

CAS No. 2070014-95-6

Selleck's AP-III-a4 (ENOblock) has been cited by 6 Publications

Purity & Quality Control

Related Other Products

Biological Activity

Description AP-III-a4 (ENOblock) is the first nonsubstrate analogue inhibitor of enolase with IC50 of 0.576 μM.
Targets
enolase [1]
(Cell-free assay)
0.576 μM
In vitro

In HCT116 cells, AP-III-a4 induces cell death under hypoxia, and inhibits cancer cell migration and invasion by down-regulation of AKT and Bcl-xL expression. In Huh7 hepatocytes and HEK kidney cells, AP-III-a4 induces glucose uptake and inhibits phosphoenolpyruvate carboxykinase (PEPCK) expression. [1]

Assay
Methods Test Index PMID
Western blot p-AMPKα / AMPKα / p-ACC / ACC ; p-AKT / AKT / p-GSK3β / GSK3β / p-PRAS40 / PRAS40 30242159
In vivo In a HCT116-xenotransplanted zebrafish tumor xenograft model, AP-III-a4 (10 μM) inhibits cancer cell migration and invasion processes. In vivo, AP-III-a4 (10 μM) also causes downregulation of PEPCK expression and induction of glucose uptake, and inhibits adipogenesis and foam cell formation. [1]

Protocol (from reference)

Kinase Assay:

[1]

  • Enolase activity assay:

    A single unit of enolase is defined as the amount of enzyme that produces 1 μmol of phosphoenol pyruvate from phospho-D-glycerate/min in standard assay. Enolase activity assay is measured at 37°C by incubating pure enolase (3–9 U) in a buffer containing 50 mM imidazole-HCl (pH 6.8), 2.0 mM MgSO4 and 400 mM KCl in the absence or presence of ENOblock or NaF. The reaction is initiated by adding 1 μmol of 2-phospho-D-glycerate, and the OD is measured after 10 min of reaction time with a spectrophometer at 240 nm.

Cell Research:

[1]

  • Cell lines: HCT116 cells
  • Concentrations: 5 μM
  • Incubation Time: 24 h
  • Method:

    3 x 105 HCT116 cells are seeded in a 6 well plate. 24 h later, cells are treated with compound of interest (with or without prior induction of hypoxia for 4 h) for 24 h using triplicate wells. Cells are then trypsinized and resuspended in 2.5 mL PBS. A 100 μL aliquot is taken for staining with 0.2% trypan blue solution and counted using a hemocytometer. 150 cells are counted and dead cells are classified as those that could not exclude trypan blue.

Animal Research:

[1]

  • Animal Models: HCT116-xenotransplanted zebrafish tumor xenograft model
  • Dosages: 10 μM
  • Administration: --

Solubility (25°C)

In vitro

Chemical Information

Molecular Weight 631.18
Formula

C31H44ClFN8O3

CAS No. 2070014-95-6
Storage 3 years -20°C powder
2 years -80°C in solvent
Smiles C1CCC(CC1)CNC2=NC(=NC(=N2)NCC3=CC=C(C=C3)F)NC4=CC=C(C=C4)CC(=O)NCCOCCOCCN.Cl

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)

mg/kg g μL

Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)

% DMSO % % Tween 80 % ddH2O
%DMSO %

Calculation results:

Working concentration: mg/ml;

Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such
as vortex, ultrasound or hot water bath can be used to aid dissolving.

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.

* Indicates a Required Field

Please enter your name.
Please enter your email. Please enter a valid email address.
Please write something to us.
Tags: buy AP-III-a4 (ENOblock) | AP-III-a4 (ENOblock) supplier | purchase AP-III-a4 (ENOblock) | AP-III-a4 (ENOblock) cost | AP-III-a4 (ENOblock) manufacturer | order AP-III-a4 (ENOblock) | AP-III-a4 (ENOblock) distributor