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Stattic STAT3 Inhibitor

Name: Stattic
Brand: Selleck Chemicals
SKU: S7024
Availability: InStock
Rating: 5 (279 reviews)

Cat.No.S7024

Stattic, the first nonpeptidic small molecule, potently inhibits STAT3 activation and nuclear translocation with IC50 of 5.1 μM in cell-free assays, highly selectivity over STAT1. Stattic induces apoptosis.

Chemical Structure

Molecular Weight: 211.19

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Quality Control

Batch: Purity: 99.88%

99.88

Related Targets	EGFR JAK Pim
Other STAT Products	Napabucasin (BBI608) NSC 74859 (S3I-201) Cryptotanshinone (Tanshinone C) SH-4-54 C188-9 (TTI-101) BP-1-102 HO-3867 Nifuroxazide AS1517499 Homoharringtonine (HHT)

Cell Culture, Treatment & Working Concentration

Cell Lines	Assay Type	Concentration	Incubation Time	Formulation	Activity Description	PMID
H9c2	Function Assay	10 μM	4 h		reverses the effects of IL-27	26339633
NPC	Function Assay	0-7.5 µM			abolishes EMT-like molecular alterations, and cell migration and invasion induced by RKIP knockdown	25915430
HASMC	Function Assay	1.25-5 μM	20 min	DMSO	inhibits p-(Y)-STAT-1,3,5 signals	25849622
H9c2	Function Assay	2/10 μM	2 h	DMSO	abrogates the cytoprotective effects of IL-27 against SH	25820907
A431	Growth Inhibition Assay	2 μM	2 h		blocks EGF-reversed decreases in cell viability	25720435
A431	Growth Inhibition Assay	2 μM	2 h		increases in apoptosis induced by shikonin	25720435
SiHa	Cell Viability Assay	5-75 nM	24 h		shows morphology of a typical apoptotic cell and dose-dependent loss of cell viability	25539644
SiHa	Function Assay	5-75 nM	24 h		reduces the phosphorylation at the tyrosine residue 705	25539644
ECA109	Growth Inhibition Assay	0-20 μM	24 h		IC50=5.50 μM	25492480
TE13	Growth Inhibition Assay	0-20 μM	24 h		IC50=6.15 μM	25492480
KYSE150	Growth Inhibition Assay	0-20 μM	24 h		IC50=12.64 μM	25492480
ECA109	Clonogenic Survival Assay	0.5 μM	24 h		suppresses the clonogenic formation	25492480
TE13	Clonogenic Survival Assay	0.5 μM	24 h		suppresses the clonogenic formation	25492480
KYSE150	Clonogenic Survival Assay	0.5 μM	24 h		suppresses the clonogenic formation	25492480
ECA109	Function Assay	0.5 μM	24 h		enhances IR-induced generation of DSBs	25492480
PC3M-1E8	Function Assay	2.5/5/10 μM	0-4 h		inhibits the STAT3 activation in a dose- and time-dependent manner	25261365
PC3M-1E8	Function Assay	10 μM	24 h		downregulates Bcl-xL, survivin and c-Myc	25261365
PC3M-1E8	Function Assay	10 μM	24 h		inhibits IL-6 induced STAT3 activation and the IL-6-induced STAT3 activation	25261365
PC3M-1E8	Clonogenic Survival Assay	2.5/5/10 μM			inhibits the colony formation significantly	25261365
MDA-MB-231	Function Assay	20 μM	2 h		exhibits Snail and E-cadherin expression	25153349
H9c2	Function Assay	20 µM	30 min	DMSO	abolishes propofol-induced AKT phosphorylation at both ser473 and thr308	25105067
HaCaT	Growth Inhibition Assay	10 µM	20 min	DMSO	enhances sorafenib- and sunitinib-induced growth inhibition	25013907
Caki-1	Growth Inhibition Assay	10 µM	20 min	DMSO	enhances sorafenib- and sunitinib-induced growth inhibition	25013907
HaCaT	Apoptosis Assay	10 µM	20 min	DMSO	increases proportions of apoptotic cells due to treatment with sorafenib or sunitinib	25013907
FHL-primed hNSCs	Cell Viability Assay	0.02-5 μM	72 h		leads to the loss of cell viability at high concentration	24945434
ELL-primed hNSCs	Cell Viability Assay	0.02-5 μM	72 h		leads to the loss of cell viability at high concentration	24945434
SS	Cell Viability Assay	1-10 μM	72 h	DMSO	causes a dose-dependent inhibition of the viability	24756111
SeAx	Cell Viability Assay	1-10 μM	72 h	DMSO	causes a dose-dependent inhibition of the viability	24756111
HuT-78	Cell Viability Assay	1-10 μM	72 h	DMSO	causes a dose-dependent inhibition of the viability	24756111
CD4+	Apoptosis Assay	10 μm	24 h	DMSO	induces apoptosis strongly	24756111
MCF-7	Growth Inhibition Assay	0.469-3.75 μM	5 d		reduces cell number significantly	24728078
MCF-7/LCC1	Growth Inhibition Assay	0.469-3.75 μM	5 d		reduces cell number significantly	24728078
MCF-7/LCC9	Growth Inhibition Assay	0.469-3.75 μM	5 d		reduces cell number significantly	24728078
HaCaT	Growth Inhibition Assay	10 µM	20 min	DMSO	enhances everolimus-induced cell growth inhibition	24423131
HaCaT	Apoptosis Assay	10 µM	20 min	DMSO	enhances the apoptotic effects of everolimus	24423131
MDA-MB-231	Function Assay	10 µM	24 h	DMSO	reduces P-STAT3 expression	24376586
SUM-159	Function Assay	10 µM	24 h	DMSO	reduces P-STAT3 expression	24376586
SK-BR-3	Function Assay	10 µM	24 h	DMSO	reduces P-STAT3 expression	24376586
MCF7-HER2	Growth Inhibition Assay	0-10 μM	48 h	DMSO	induces cell death dose dependently	24297508
MCF7-HER2	Function Assay	5 μM	24 h	DMSO	diminishes Sox-2, Oct-4, and slug expression	24297508
MCF7-HER2	Function Assay	5 μM	24 h	DMSO	decreases the expression levels of EMT markers, vimentin and slug	24297508
MCF7-HER2	Growth Inhibition Assay	5 μM	24 h	DMSO	enhances cell growth inhibition combined with Herceptin	24297508
HMECs	Function Assay	10 μM	2 h		inhibits IFNα mediated phosphorylation of STAT1, STAT2 and STAT3	24211327
HTR8/SVneo	Function Assay	1 μM	1 h		suppressed OSM-induced STAT3 phosphorylation	24060241
HTR8/SVneo	Function Assay	0.5/1 μM	48 h		restores the expression of E-cadherin suppressed by OSM	24060241
HTR8/SVneo	Function Assay	1 μM	48 h		significantly increases migration by OSM	24060241
C13*	Apoptosis Assay	0-10 μM	24/48 h		induces apoptosis in a dose and time dependent manner	23962558
OV2008	Apoptosis Assay	0-10 μM	24/48 h		induces apoptosis in a dose and time dependent manner	23962558
C13*	Apoptosis Assay		24/48 h		enhances cisplatin-induced apoptosis	23962558
OV2008	Apoptosis Assay		24/48 h		enhances cisplatin-induced apoptosis	23962558
W480	Function Assay	2.5/10 μM	30 min	DMSO	sensitizes cells to chemoradiotherapy in a dose-dependent manner	23934972
SW837	Function Assay	2.5/10 μM	30 min	DMSO	sensitizes cells to chemoradiotherapy in a dose-dependent manner	23934972
T24	Function Assay	2/10/20 μM	24 h		causes dose-dependent inhibition of the CXCL12-induced increase of invading cells	23526079
CNE1	Function Assay	20 µM	48 h		blocks the IL-6 increased phosphorylation of Stat3	23382914
CNE2	Function Assay	20 µM	48 h		blocks the IL-6 increased phosphorylation of Stat3	23382914
HONE1	Function Assay	20 µM	48 h		blocks the IL-6 increased phosphorylation of Stat3	23382914
CNE1	Growth Inhibition Assay	4 μM			significantly reduces cell viability	23382914
CNE1	Function Assay	0-20 μM	0-4 h		inhibits Stat3 activation in a dose- and time-dependent manner	23382914
CNE2	Function Assay	0-20 μM	0-4 h		inhibits Stat3 activation in a dose- and time-dependent manner	23382914
HONE1	Function Assay	0-20 μM	0-4 h		inhibits Stat3 activation in a dose- and time-dependent manner	23382914
CNE1	Cell Viability Assay	0.5-64 μM	48 h		suppresses cell viability in a dose- and time-dependent manner	23382914
CNE2	Cell Viability Assay	0.5-64 μM	48 h		suppresses cell viability in a dose- and time-dependent manner	23382914
HONE1	Cell Viability Assay	0.5-64 μM	48 h		suppresses cell viability in a dose- and time-dependent manner	23382914
C666-1	Cell Viability Assay	0.5-64 μM	48 h		suppresses cell viability in a dose- and time-dependent manner	23382914
CNE1	Apoptosis Assay	10 µM	48 h		induces apoptosis	23382914
CNE2	Apoptosis Assay	10 µM	48 h		induces apoptosis	23382914
HONE1	Apoptosis Assay	10 µM	48 h		induces apoptosis	23382914
CNE2	Cell Viability Assay	1/2 μM	48 h		sensitize cells to radiotherapy	23382914
HONE1	Cell Viability Assay	1/2 μM	48 h		sensitize cells to radiotherapy	23382914
C666-1	Cell Viability Assay	1/2 μM	48 h		sensitize cells to radiotherapy	23382914
HEC-1A	Function Assay	1 μM	24 h	DMSO	blocks the MUC20-enhanced invasion triggered by 10% FBS	23262208
RL95-2	Function Assay	1 μM	24 h	DMSO	blocks the MUC20-enhanced invasion triggered by 10% FBS	23262208
HEC-1A	Function Assay	1 μM	24 h	DMSO	blocks the MUC20-enhanced invasion triggered by EGF	23262208
RL95-2	Function Assay	1 μM	24 h	DMSO	blocks the MUC20-enhanced invasion triggered by EGF	23262208
CT26	Function Assay	20 mM	1 h		suppresses HGF-induced VEGF expression	23233163
UM-SCC-17B	Growth Inhibition Assay				IC50=2.562 ± 0.409 μM, GI50=1.279 ± 0.194 μM	22770899
OSC-19	Growth Inhibition Assay				IC50=3.481 ± 0.953 μM, GI50=1.366 ± 0.770 μM	22770899
Cal33	Growth Inhibition Assay				IC50=2.282 ± 0.423 μM, GI50=1.349 ± 0.363 μM	22770899
UM-SCC-22B	Growth Inhibition Assay				IC50=2.648 ± 0.542 μM, GI50=1.320 ± 0.204 μM	22770899
UM-SCC-17B	Function Assay	0-30 μM	0-24 h		inhibits STAT3 activation dose and time dependently	22770899
OSC-19	Function Assay	0-30 μM	0-24 h		inhibits STAT3 activation dose and time dependently	22770899
Cal33	Function Assay	0-30 μM	0-24 h		inhibits STAT3 activation dose and time dependently	22770899
UM-SCC-22B	Function Assay	0-30 μM	0-24 h		inhibits STAT3 activation dose and time dependently	22770899
U-87MG	Cell Viability Assay	0-10 μM	72 h	DMSO	inhibits cell viability dose dependently	25436682
U-373MG	Cell Viability Assay	0-10 μM	72 h	DMSO	inhibits cell viability dose dependently	25436682
SH-SY5Y	Cell Viability Assay	0-10 μM	72 h	DMSO	inhibits cell viability dose dependently	25436682
Tu-9648	Cell Viability Assay	0-10 μM	72 h	DMSO	inhibits cell viability dose dependently	25436682
Neuro-2a	Cell Viability Assay	0-10 μM	72 h	DMSO	inhibits cell viability dose dependently	25436682
PCNs	Cell Viability Assay	0-10 μM	72 h	DMSO	inhibits cell viability dose dependently	25436682
PGCs	Cell Viability Assay	0-10 μM	72 h	DMSO	inhibits cell viability dose dependently	25436682
RAW264.7	Function Assay	10 μM	12 h		abrogates the mRNA expressions of JAK2, STAT1, STAT2, and STAT3 induced by DON and T-2 toxin	22454431
RAW264.7	Apoptosis Assay	5/10 μM	45 min		enhances toxins induced apoptosis and MMP loss	22454431
SW480	Cell Viability Assay	5/10/20 μM	72 h		inhibits cell viability of the ALDH+/CD133+ cells	21900397
HCT116	Cell Viability Assay	5/10/20 μM	72 h		inhibits cell viability of the ALDH+/CD133+ cells	21900397
DLD-1	Cell Viability Assay	5/10/20 μM	72 h		inhibits cell viability of the ALDH+/CD133+ cells	21900397
SNU387	Cell Viability Assay	20 μM	24 h		reduces cell viability	21311975
SNU398	Cell Viability Assay	20 μM	24 h		reduces cell viability	21311975
HepG2	Cell Viability Assay	20 μM	24 h		reduces cell viability	21311975
Huh-7	Cell Viability Assay	20 μM	24 h		reduces cell viability	21311975
VSMC	Growth Inhibition Assay	3/5/10 μM	30 min	DMSO	prevents PDGF- and thrombin-mediated VSMC proliferation in a dose-dependent manner	20847306
MDA-MB-231	Apoptosis Assay	10 μM	24 h	DMSO	induces apoptosis	17114005
MDA-MB-435S	Apoptosis Assay	10 μM	24 h	DMSO	induces apoptosis	17114005
AsPC1	Antiproliferative assay		72 hrs		Antiproliferative activity against human AsPC1 cells assessed as inhibition of cell proliferation after 72 hrs by MTS assay, IC50 = 1.32 μM.	24904966
MDA-MB-231	Antiproliferative assay		72 hrs		Antiproliferative activity against ER-negative and triple-negative human MDA-MB-231 cells assessed as inhibition of cell proliferation after 72 hrs by MTS assay, IC50 = 2.89 μM.	24904966
MCF7	Antiproliferative assay		72 hrs		Antiproliferative activity against ER-positive human MCF7 cells assessed as inhibition of cell proliferation after 72 hrs by MTS assay, IC50 = 3.6 μM.	24904966
PANC1	Antiproliferative assay		72 hrs		Antiproliferative activity against human PANC1 cells assessed as inhibition of cell proliferation after 72 hrs by MTS assay, IC50 = 3.77 μM.	24904966
MDA-MB-231	Cytotoxicity assay		48 hrs		Cytotoxicity against human MDA-MB-231 cells assessed as growth inhibition after 48 hrs by MTT assay, IC50 = 1.56 μM.	26396689
MDA-MB-435S	Cytotoxicity assay		48 hrs		Cytotoxicity against human MDA-MB-435S cells assessed as growth inhibition after 48 hrs by MTT assay, IC50 = 1.87 μM.	26396689
MCF7	Cytotoxicity assay		48 hrs		Cytotoxicity against human MCF7 cells assessed as growth inhibition after 48 hrs by MTT assay, IC50 = 2.16 μM.	26396689
A549	Cytotoxicity assay		48 hrs		Cytotoxicity against human A549 cells assessed as growth inhibition after 48 hrs by MTT assay, IC50 = 2.5 μM.	26396689
DU145	Cytotoxicity assay		48 hrs		Cytotoxicity against human DU145 cells assessed as growth inhibition after 48 hrs by MTT assay, IC50 = 2.5 μM.	26396689
PANC1	Cytotoxicity assay		48 hrs		Cytotoxicity against human PANC1 cells assessed as growth inhibition after 48 hrs by MTT assay, IC50 = 2.9 μM.	26396689
HCT116	Antiproliferative assay		72 hrs		Antiproliferative activity against human HCT116 cells after 72 hrs by MTT assay, IC50 = 1.08 μM.	27718470
MDA-MB-231	Antiproliferative assay		72 hrs		Antiproliferative activity against human MDA-MB-231 cells after 72 hrs by MTT assay, IC50 = 1.68 μM.	27718470
MCF7	Antiproliferative assay		72 hrs		Antiproliferative activity against human MCF7 cells after 72 hrs by MTT assay, IC50 = 2.36 μM.	27718470
A549	Antiproliferative assay		72 hrs		Antiproliferative activity against human A549 cells after 72 hrs by MTT assay, IC50 = 4.4 μM.	27718470
AD293	Function assay		6 hrs		Inhibition of IFNgamma-stimulated GFP/FLAG-tagged STAT3 dimerization in human AD293 cells incubated for 6 hrs by Western blot analysis, IC50 = 5.1 μM.	30228000
MDA-MB-231	Function assay	1 to 10 uM	12 hrs		Inhibition of STAT3 phosphorylation at Tyr705 in human MDA-MB-231 cells at 1 to 10 uM after 12 hrs by western blot analysis	24904966
MDA-MB-231	Anticancer assay	1 to 10 uM	48 hrs		Anticancer activity against human MDA-MB-231 cells assessed as cell growth inhibition, apoptosis and cellular morphological changes at 1 to 10 uM after 48 hrs by light microscopy	24904966
MDA-MB-231	Function assay	1 to 10 uM	12 hrs		Decrease in STAT3 protein expression in human MDA-MB-231 cells at 1 to 10 uM after 12 hrs by western blot analysis	24904966
MCF7	Function assay		12 hrs		Inhibition of STAT3 phosphorylation at Y705 in human MCF7 cells after 12 hrs by Western blot analysis	26396689
MDA-MB-435S	Function assay		12 hrs		Inhibition of STAT3 phosphorylation at Y705 in human MDA-MB-435S cells after 12 hrs by Western blot analysis	26396689
MDA-MB-231	Function assay		12 hrs		Inhibition of STAT3 phosphorylation at Y705 in human MDA-MB-231 cells after 12 hrs by Western blot analysis	26396689
Click to View More Cell Line Experimental Data

Solubility

In vitro
Batch:

DMSO : 42 mg/mL (198.87 mM)
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Water : Insoluble

Ethanol : Insoluble

Molarity Calculator

Mass	Concentration	Volume	Molecular Weight
Mass value Mass unit	Concentration value Concentration unit	Volume value Volume unit	Molecular weight (g/mol)

Dilution Calculator Molecular Weight Calculator

In vivo batch selection In vivo Batch:
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution	5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	2.100mg/ml (9.94mM)	Taking the 1 mL working solution as an example, add 50 μL of 42 mg/ml clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to clarify; then continue to add 500 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution	5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	1.000mg/ml (4.74mM)	Taking the 1 mL working solution as an example, add 50 μL of 10.9 mg/ml clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to clarify; then continue to add 500 μL of ddH2O to make it clear. Volume up to 1 mL. The mixed solution should be used immediately for optimal results.
Clear solution	5% DMSO 1 95% Corn oil Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	0.800mg/ml (3.79mM)	Taking the 1 mL working solution as an example, add 50 μL of 16 mg/ml clear DMSO stock solution to 950 μL of corn oil and mix evenly. The mixed solution should be used immediately for optimal results.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution	10%DMSO 1 40%PEG300 2 5%Tween80 3 45%ddH2O Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	4.800mg/ml (22.73mM)	Taking the 1 mL working solution as an example, add 100 μL of 48 mg/ml clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to make it clear; then continue to add 450 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.

In vivo Formulation Calculator (Clear solution)

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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.

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Chemical Information, Storage & Stability

Molecular Weight	211.19	Formula	C₈H₅NO₄S	Storage (From the date of receipt)	3 years-20°Cpowder
CAS No.	19983-44-9	Download SDF		Storage of Stock Solutions	1 year-80°Cin solvent 1 month-20°Cin solvent
Synonyms	N/A			Smiles	C1=CC(=CC2=C1C=CS2(=O)=O)[N+](=O)[O-]

Mechanism of Action

Features	Stattic is the first non-peptide small molecule with inhibitory activity against STAT3 SH2 domain regardless of the STAT3 phosphorylation state in vitro.
Targets/IC₅₀/Ki	STAT3 (Cell-free assay) 5.1 μM
In vitro	Stattic inhibits binding of a phosphotyrosine-containing peptide derived from the gp130 receptor to the STAT3 SH2 domain in a strongly temperature-dependent manner. This compound only has a very weak effect on binding of a tyrosinephosphorylated peptide to the SH2-domain of the tyrosine kinase Lck. And it doesn’t inhibit dimerization of two other dimeric transcription factors (c-Myc/Max and Jun/Jun). It also inhibits fluorescein-labeled phosphopeptides to the SH2 domains of STAT1 and STAT5b. This chemical selectively inhibits DNA binding of STAT3 homodimers at a concentration of 10 μM. Shown to inhibit cellular phosphorylation of STAT3 at Tyr705 with little effect towards STAT1 phosphorylation at Tyr701 (in HepG2 cells) or the phosphorylation of JAK1, JAK2, and c-Src (in MDA-MB-231 and MDA-MB-235S cells). This compound increases the apoptotic rate of STAT3-dependent breast cancer cell lines.
Kinase Assay	High-Throughput Screening and FluorescencePolarization Assays
Kinase Assay	Screening is performed at approximately 30 °C. The specificity of screening hits is validated in analogous assays for binding of the test compounds to the SH2 domains of STAT1, STAT5, and Lck. The final concentration of buffer components used for all FP assays is 10 mM HEPES (pH 7.5), 1 mM EDTA, 0.1% Nonidet P-40, 50 mM NaCl, and 10% DMSO. The absence of dithiothreitol is essential for inhibitory activity. The sequences of the peptides are: STAT3, 5-carboxyfluorescein-GY(PO₃H₂)LPQTV-NH₂; STAT1, 5-carboxyfluorescein-GY(PO₃H₂)DKPHVL; STAT5, 5-carboxyfluorescein-GY(PO₃H₂)LVLDKW; and Lck, 5-carboxyfluorescein-GY(PO₃H₂)EEIP. For specificity analysis at 30 °C, proteins are used at 150 nM (STAT1, STAT3, and STAT5). For specificity analysis at 37 °C, proteins are used at 370 nM (STAT3) or 100 nM (Lck). Proteins are incubated with test compounds in Eppendorf tubes at the indicated temperatures for 60 min prior addition of the respective 5-carboxyfluorescein labeled peptides (final concentration: 10 nM). Before measurement at room temperature, the mixtures are allowed to equilibrate for at least 30 min. Test compounds are used at the indicated concentrations diluted from a 20× stock in DMSO. Binding curves and inhibition curves are fitted with SigmaPlot. All competition curves are repeated three times in independent experiment
References	[1] https://pubmed.ncbi.nlm.nih.gov/17114005/ [2] https://pubmed.ncbi.nlm.nih.gov/21521803/

Applications

Methods	Biomarkers	PMID
Western blot	PARP / C-PARP / Caspase-3 / C-Caspse-3 Survivin / c-Myc / Bcl-xl p-STAT3 / STAT3	23382914
Immunofluorescence	p-STAT3 / STAT3 / Survivin	25261365
Growth inhibition assay	Cell viability	23382914
ELISA	BDNF	27456333