1. MAPK/ERK Pathway
  2. JNK

JNK-IN-7 (Synonyms: JNK inhibitor)

Cat. No.: HY-15617 Purity: 98.39%
Data Sheet SDS Handling Instructions

JNK-IN-7 is a potent JNK inhibitor with IC50 of 1.5, 2 and 0.7 nM for JNK1, JNK2 and JNK3, respectively.

For research use only. We do not sell to patients.
JNK-IN-7 Chemical Structure

JNK-IN-7 Chemical Structure

CAS No. : 1408064-71-0

Size Price Stock Quantity
10 mM * 1 mL in DMSO $315 In-stock
5 mg $286 In-stock
10 mg $369 In-stock
50 mg $996 In-stock
100 mg $1588 In-stock
200 mg   Get quote  
500 mg   Get quote  

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  • Biological Activity

  • Protocol

  • Technical Information

  • Purity & Documentation

  • References

Description

JNK-IN-7 is a potent JNK inhibitor with IC50 of 1.5, 2 and 0.7 nM for JNK1, JNK2 and JNK3, respectively.

IC50 & Target

IC50: 1.5 nM (JNK1), 2 nM (JNK2), 0.7 nM (JNK3)[1]

In Vitro

JNK-IN-7 is a relatively selective JNK inhibitor in cells. In addition to JNK 1, 2, 3, JNK-IN-7 also binds to IRAK1(IC50=14.1 nM), YSK4 (IC50=4.8 nM), ERK3 (IC50=22 nM), PIK3C3, PIP5K3 and PIP4K2C[1]. Expression of divalent metal-ion transporter 1 (DMT1) in HCT116 is demonstrated to be markedly decreased under stimulation with TNF for 24 and 48 h, while JNK-IN-7 can significantly reverse the decrease. TNF can down-regulate DMT1 expression, while JNK-IN-7 can markedly suppress this function[2].

References
Preparing Stock Solutions
Concentration Volume (DMSO) Mass 1 mg 5 mg 10 mg
1 mM 2.0261 mL 10.1305 mL 20.2610 mL
5 mM 0.4052 mL 2.0261 mL 4.0522 mL
10 mM 0.2026 mL 1.0130 mL 2.0261 mL
Kinase Assay
[1]

A375 cells are pre-treated with 1μM JNK-IN-7 for the indicated amounts of time. Remove the medium and wash 3 times with PBS. Resuspend the cell pellet with 1 mL Lysis Buffer (1% NP-40, 1% CHAPS, 25 mM Tris, 150 mM NaCl, Phosphatase Inhibitor Cocktail). Rotate end-to-end for 30 min at 4°C. Lysates are cleared by centrifugation at 14000 rpm for 15 min in the Eppendorf. The cleared lysates gel filtered into Kinase Buffer (0.1% NP-40, 20 mM HEPES, 150 mM NaCl, Phosphatase Inhibitor Cocktail, Protease Inhibitor Cocktail) using Bio-Rad 10DG colums. The total protein concentration of the gel-filtered lysate should be around 5-15 mg/mL. Cell lysate is labeled with the probe at 5 μM for 1 hour. Samples are reduced with DTT, and cysteines are blocked with iodoacetamide and gel filtered to remove excess reagents and exchange the buffer. Add 1 volume of 2X Binding Buffer (2% Triton-100, 1% NP-40, 2 mM EDTA, 2X PBS) and 50 μL streptavidin bead slurry and rotate end-to-end for 2 hours, centrifuge at 7000 rpm for 2 min. Wash 3 times with 1X Binding Buffer and 3 times with PBS. Add 30 μL 1X sample buffer to beads, heat samples at 95°C for 10 min. Run samples on an SDS-PAGE gel at 110V. After transferred, the membrane is immunoblotted with JNK antibody[1]. MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Cell Assay
[2]

JNK-IN-7 is prepared in DMSO and stored, and then diluted with appropriate medium before use[2].

Intestinal epithelial cell line (HCT116) is cultured in DMEM medium, supplemented with 10% heat-inactivated fetal bovine serum (FBS), penicillin (100 U/mL) and streptomycin (100 g/mL), 2 mM L-gentamycin, and 50 μM 2-ME. These cells are stimulated with TNF (20 ng/mL), LPS (100 ng/mL), and IFN-γ (20 ng/mL), respectively. After 24 or 48 h of culture, cells are harvested followed by extraction of total RNA, and the levels of DMT1 mRNA are analyzed by qRT-PCR. To determine the mechanisms of TNF involved in regulating DMT1 expression, JNK-IN-7 (1 μM), NF-κB inhibitor (BAY 11-7082, 1 μM), and caspase-3/8 inhibitor (Z-DEVD-FMK, 50 μM) are also added into the culture medium. After 48 h of culture, cells are then collected to detect the expression of DMT1 by qRT-PCR[2]. MCE has not independently confirmed the accuracy of these methods. They are for reference only.

References
Molecular Weight

493.56

Formula

C₂₈H₂₇N₇O₂

CAS No.

1408064-71-0

Storage

Please store the product under the recommended conditions in the Certificate of Analysis.

Shipping

Room temperature in continental US; may vary elsewhere

Solvent & Solubility

10 mM in DMSO

* "<1 mg/mL" means slightly soluble or insoluble. "≥" means soluble, but saturation unknown.

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Product Name:
JNK-IN-7
Cat. No.:
HY-15617
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