HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：11812 靶标：N/A

External ID: FATTTLab-Algae-Lipid

Protocol: Cells were pre-grown to mid-log phase in TAP media. For primary screen, cells were seeded at low density of 500,000 / well in 384-well plate. Compound in DMSO was added to each well at a final concentration of 10 micromolar and cells were allowed to grow for 72 hours. At the end of assay, Nile Red (30 uM f.c.) was added and plates were incubated at 37 C for 60 mins in dark. Fluorescence intensity was measured. Readouts were reported as normalized fold change in the intensity of treated versus control.

Comment: Reference:
1. Wase, N., Tu, B., Black, P. N. & DiRusso, C. C. Phenotypic screening identifies Brefeldin A/Ascotoxin as an inducer of lipid storage in the algae Chlamydomonas reinhardtii. Algal Research 11, 74-84 (2015).

2. Identification and metabolite profiling of chemical activators of lipid accumulation in green algae
#Nishikant Wase, Boqiang Tu, James W Allen, Paul N Black, Concetta C DiRusso
#Plant Physiology Jun 2017, pp.00433.2017; DOI: 10.1104/pp.17.00433
#http://www.plantphysiol.org/content/early/2017/06/26/pp.17.00433

Patent:
DiRusso, C., & Wase, N. (2016). Compounds for Increasing Lipid Synthesis and Storage. United States#NUtech Ventures (Lincoln, NE, US) http://www.freepatentsonline.com/y2016/0312253.html

For additional Information please contact:

Prof. Concetta C. DiRusso
Department of Biochemistry
University of Nebraska-Lincoln
cdirusso2@unl.edu

Nishikant Wase, PhD
Department of Biochemistry
University of Nebraska-Lincoln
nishikant.wase@gmail.com

HIT_COMPOUND_IN_SCREEN
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：Hemozoin

External ID: CHEMBL4428589

Protocol: N/A

Comment: Compounds with activity <= 10uM or explicitly reported as active by ChEMBL are flagged as active in this PubChem assay presentation.

Journal: J Med Chem
Year: 2016
Volume: 59
Issue: 13
First Page: 6512
Last Page: 6530
DOI: 10.1021/acs.jmedchem.6b00719

Target ChEMBL ID: CHEMBL613898
ChEMBL Target Name: Hemozoin
ChEMBL Target Type: MACROMOLECULE - Target is a biological macromolecule (e.g., glycoproteins, hemozoin, hydroxyapatite)
Relationship Type: M - Molecular target other than protein assigned
Confidence: Target assigned is molecular non-protein target

PubChem Standard Value	Standard Type	Standard Relation	Standard Value	Standard Units	Data Validity Comment
4.3	IC50	=	4300	nM
22	IC50	=	22000	nM
8	IC50	=	8000	nM
21	IC50	=	21000	nM
5500	IC50	>	5500000	nM	Outside typical range
5500	IC50	>	5500000	nM	Outside typical range
5500	IC50	>	5500000	nM	Outside typical range
5500	IC50	>	5500000	nM	Outside typical range
10.9	IC50	=	10900	nM
7	IC50	=	7000	nM
260	IC50	=	260000	nM	Outside typical range
134	IC50	=	134000	nM	Outside typical range

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：24983 靶标：Huntingtin

External ID: KUHTS-Muma KU-CaM-Htt INH-01

Protocol: 1; Dispense 45 nl compounds (10 mM stocks) using ECHO 555 to Alpha 384 well assay plates. Dispense 45 nl DMSO to control columns 1 and 2 of 384 well plates.
2; Incubate 5 ul of 6XHis-mHTT (Final, 13 nM) with the compounds for 40 mins at room temperature in buffer containing 10 mM Tris.HCl pH 7.4, 1 mM calcium chloride, 150 mM sodium chloride, 0.1% BSA and 20% glycerol.
3; Dispense 5 ul of 6XGST-CaM (Final 13 nM) in buffer A.
4; Incubation; 1 hour (dark at 25C)
5; Dispense 20 ul of Nickel chelate acceptor beads (Final, 20 ugs/ml) and Glutathione donor beads (Final, 30 ugs/ml). Incubate for 2h, room temperature.
6; Detector: Perkin Elmer Enspire, Alphascreen Module (Excitation 680nm/Emission 570nm).

NOTES (numbers refer to Sequence numbers above)
1. Alphascreen bead incubations were performed in green light, TiterTek setting 400rpm.
2. All incubation and addition steps were followed by mixing and centrifugation at 400g,1 min.
3. The percent inhibition for each compound was calculated as follows:
100- [100 *((Test Compound-Median Low Control) / (Median High Control - Median Low Control))]

Where:
Test_Compound is defined as wells containing His mHTT + GST CaM in the presence of test compound

High_Control is defined as wells containing His mHTT + GST CaM and DMSO.

Low_Control is defined as wells containing His mHTT and DMSO.

Comment: All percent inhibition data reported were normalized to high and low controls on a per-plate basis. The results of primary screening data include compounds contributing to assay signal interference, interference with tags binding to Alpha beads, chelators, process related artifacts etc.. The actives were defined as compounds that inhibited Alphascreen reads to greater than 50%.

Activity at 15 uM	Phenotype
-3.1
4.2
-4.8
-0.6
-3.9
-6
75.3	Inhibitor
-10.3
-7
-2
-3.8
17.6
-4.8
-4.1
-7.8
-7.5
15.2
7.8
-2.5
-13.4

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：Ferriprotoporphyrin IX

External ID: CHEMBL4428622

Protocol: N/A

Comment: Journal: J Med Chem
Year: 2016
Volume: 59
Issue: 13
First Page: 6512
Last Page: 6530
DOI: 10.1021/acs.jmedchem.6b00719

Target ChEMBL ID: CHEMBL613897
ChEMBL Target Name: Ferriprotoporphyrin IX
ChEMBL Target Type: SMALL MOLECULE - Target is a small molecule such as an amino acid, sugar or metabolite)
Relationship Type: M - Molecular target other than protein assigned
Confidence: Target assigned is molecular non-protein target

Standard Type	Activity Comment
Activity	Not Active
Activity	Not Active
Activity	Not Active
Activity	Not Active
Activity	Not Active

14547-75-2 靶点实验数据