JNK-IN-8
Based on 59 publication(s) in Google Scholar
JNK-IN-8 (JNK Inhibitor XVI) is a potent JNK inhibitor with IC50s of 4.7 nM, 18.7 nM, and 1 nM for JNK1, JNK2, and JNK3, respectively.
For research use only. We do not sell to patients.
- Purity: 99.67%
- CAS No.: 1410880-22-6
- Formula: C29H29N7O2
- Molecular Weight:507.59
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Storage:Powder -20°C, 3 years , 4°C, 2 years ; In solvent -80°C, 2 years , -20°C, 1 year
Publications Citing Use of MedChemExpress (MCE) JNK-IN-8
More- Nat Nanotechnol. 2021 Jul;16(7):830-839. [Abstract]
- Mil Med Res. 2023 Jun 5;10(1):25. [Abstract]
- Nat Commun. 2026 Feb 12;17(1):1214. [Abstract]
- Nat Commun. 2025 Feb 7;16(1):1429. [Abstract]
- Nat Commun. 2024 Nov 21;15(1):10106. [Abstract]
- Nat Commun. 2025 Jan 9;16(1):515. [Abstract]
- Nat Commun. 2020 Jan 3;11(1):71. [Abstract]
- Cell Death Differ. 2020 May;27(5):1569-1587. [Abstract]
- Adv Sci (Weinh). 2025 Aug 4:e03972. [Abstract]
- J Clin Invest. 2026 Mar 17:e199056. [Abstract]
- J Clin Invest. 2024 Jan 9;134(4):e174984. [Abstract]
- J Exp Clin Cancer Res. 2018 May 4;37(1):99. [Abstract]
- Cell Discov. 2026 Feb 24;12(1):13. [Abstract]
- Cell Rep Med. 2026 Apr 21;7(4):102691. [Abstract]
- Cell Rep Med. 2025 Apr 25:102098. [Abstract]
- J Neuroinflammation. 2025 Oct 28;22(1):246. [Abstract]
- Acta Biomater. 2021 Nov;135:567-581. [Abstract]
- Proc Natl Acad Sci U S A. 2023 Dec 12;120(50):e2313148120. [Abstract]
- Dev Cell. 2021 Dec 20;56(24):3334-3348.e6. [Abstract]
- Cell Syst. 2022 Oct 30;S2405-4712(22)00430-6. [Abstract]
- J Transl Med. 2025 Jun 3;23(1):621. [Abstract]
- Cell Rep. 2021 Oct 12;37(2):109806. [Abstract]
- Clin Transl Med. 2022 Feb;12(2):e747. [Abstract]
- Sci Signal. 2026 Jan 13;19(920):eadw3709. [Abstract]
- Sci Signal. 2025 Aug 12;18(899):eads7002. [Abstract]
- J Ethnopharmacol. 2024 Jan 30;319(Pt 2):117270. [Abstract]
- Cells. 2022 Apr 25;11(9):1451. [Abstract]
- Life Sci. 2025 May 11:123698. [Abstract]
- Commun Biol. 2021 Mar 25;4(1):399. [Abstract]
- Cancer Gene Ther. 2026 May 2. [Abstract]
- Int J Mol Sci. 2022 Sep 20;23(19):11004. [Abstract]
- Expert Opin Ther Targets. 2024 May;28(5):461-471. [Abstract]
- J Dairy Sci. 2020 Dec;103(12):11636-11652. [Abstract]
- Poult Sci. 2025 Dec 5;105(1):106210. [Abstract]
- J Cell Mol Med. 2025 Jun;29(11):e70624. [Abstract]
- J Cell Mol Med. 2022 Aug;26(16):4602-4612. [Abstract]
- Toxicol Sci. 2026 Jun 5;209(6):kfag060. [Abstract]
- J Biol Chem. 2014 Oct 17;289(42):28753-64. [Abstract]
- Cell Signal. 2023 May:105:110607. [Abstract]
- Mol Med Rep. 2021 Feb;23(2):150. [Abstract]
- BMC Cancer. 2020 Mar 12;20(1):214. [Abstract]
- Toxicol Appl Pharmacol. 2019 Apr 1:368:37-48. [Abstract]
- Oral Dis. 2025 Oct 27. [Abstract]
- Mol Cell Biol. 2021 Feb 23;41(3):e0033320. [Abstract]
- Reprod Sci. 2025 Dec 29. [Abstract]
- Thorac Cancer. 2021 May;12(10):1558-1569. [Abstract]
- Leuk Lymphoma. 2021 Dec;62(14):3361-3372. [Abstract]
- Appl Bionics Biomech. 2022 Aug 16:2022:8460121. [Abstract]
- Npj Viruses. 2025 Sep 18;3(1):69. [Abstract]
- University of Minnesota. 2025.
- bioRxiv. 2025 Feb 9:2025.02.08.637251. [Abstract]
- Res Sq. 2024 Jul 31.
- bioRxiv. 2024 September 14.
- bioRxiv. 2024 July 12.
- University of Gothenburg. 2023 Jun 27.
- Research Square Preprint. 2021 Feb.
- bioRxiv. 2020 Apr.
- Harvard University. 2019 May.
- Harvard Medical School LINCS LIBRARY
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In Vivo Efficacy Study
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RT-PCR
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Flow Cytometry
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Flow Cytometry
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IF
Biological Activity
|
JNK3 1 nM (IC50) |
JNK1 4.7 nM (IC50) |
JNK2 18.7 nM (IC50) |
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Cell Line
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Type | Value | Description | References |
|---|---|---|---|---|
| A-375 | IC50 |
0.338 μM
Compound: 72
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Inhibition of JNK3-mediated c-jun phosphorylation in human A375 cells after 1 hr incubation
Inhibition of JNK3-mediated c-jun phosphorylation in human A375 cells after 1 hr incubation
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[PMID: 25415535] |
| HeLa | IC50 |
0.486 μM
Compound: 72
|
Inhibition of JNK3-mediated c-jun phosphorylation in human HeLa cells after 1 hr incubation
Inhibition of JNK3-mediated c-jun phosphorylation in human HeLa cells after 1 hr incubation
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[PMID: 25415535] |
JNK-IN-8 inhibits phosphorylation of c-Jun, a direct substrate of JNK kinase. JNK-IN-8 inhibits c-Jun phosphorylation in HeLa and A375 cells with EC50 of 486 nM and 338 nM, respectively. JNK-IN-8 also exhibits exceptional selectivity based upon KinomeScan and enzymatic profiling. Cumulatively these combined profiling technologies demonstrate that both JNK-IN-8 and JNK-IN-12 are remarkably selective covalent JNK inhibitors and are appropriate for interrogating JNK-dependent biological phenomena[1].JNK-IN-8, a selective pan-JNK inhibitor, is discovered to inhibit JNK kinase by broad-base kinase selectivity profiling of a library of acrylamide kinase inhibitors based on the structure of imatinib using the KinomeSca approach. JNK-IN-8 possess distinct regio-chemistry of the 1,4-dianiline and 1,3-aminobenzoic acid substructures relative to imatinib and uses an N,N-dimethyl butenoic actemide warhead to covalently target Cys154. JNK-IN-8 adopts an L-shaped type I binding conformation to access Cys 154 located towards the lip of the ATP-binding site[2].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
Chemical Information
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CAS No. 1410880-22-6
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Appearance Solid
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Molecular Weight 507.59
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Formula C29H29N7O2
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Color White to yellow
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SMILES
CC1=CC(NC(C2=CC=CC(NC(/C=C/CN(C)C)=O)=C2)=O)=CC=C1NC3=NC=CC(C4=CN=CC=C4)=N3
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Synonyms
JNK Inhibitor XVI
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Shipping
Room temperature in continental US; may vary elsewhere.
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Storage
Powder -20°C 3 years 4°C 2 years In solvent -80°C 2 years -20°C 1 year
Publications (59)
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Journal Impact Factor
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Most Recent
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Nat Nanotechnol
Therapeutically reprogrammed nutrient signalling enhances nanoparticulate albumin bound drug uptake and efficacy in KRAS-mutant cancer. [Abstract]2021 Jul;16(7):830-839. PMID: 33958764 -
Mil Med Res
2023 Jun 5;10(1):25. PMID: 37271807
JNK-IN-8 purchased from MedChemExpress. Usage Cited in: Mil Med Res. 2023 Jun 5;10(1):25. [Abstract]
The proliferation ability of PC-3 cells transfected with shASPA and/or treated with JNK-IN-8 (2 μM; 0-5 d) was assessed using a CCK-8 assay.
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Nat Commun
Human iPSC-based Modeling of Pulmonary Fibrosis Reveals p300/CBP Inhibition Suppresses Alveolar Transitional Cell State. [Abstract]2026 Feb 12;17(1):1214. PMID: 41680175 -
Nat Commun
2025 Feb 7;16(1):1429. PMID: 39920102
JNK-IN-8 purchased from MedChemExpress. Usage Cited in: Nat Commun. 2025 Feb 7;16(1):1429. [Abstract]
g-j. The effects of JNK (JNK-IN-8) (25 nM) and PKC (Go 6983) (100 nM) inhibition on the expression of Cxcl10 and Ccl4 in MC38 and Panc02 cells.
JNK-IN-8 purchased from MedChemExpress. Usage Cited in: Nat Commun. 2025 Feb 7;16(1):1429. [Abstract]
FACS analysis of CD8+ T-cell proliferation (CFSE low) in cultured with conditioned media derived from MC38 cells treated with WNT11 and inhibitors of CAMKII (CAMKII-IN-1) (100 nM), JNK (JNK-IN-8) (25 nM) and PKC (Go 6983) (100 nM) as indicated.
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Nat Commun
2024 Nov 21;15(1):10106. PMID: 39572544
JNK-IN-8 purchased from MedChemExpress. Usage Cited in: Nat Commun. 2024 Nov 21;15(1):10106. [Abstract]
p, q. Mice were treated with JNK-IN-8 (10 mg/kg, i.p., every other day for 15 days) inhibits NF2 LOF mutants-induced IOMM-Lee xenograft tumor growth. The collected tumors treated with DMSO or JNK-IN-8 were shown (p) and tumor weight was measured and quantified (mean ± s.d., n = 6 mice per group) (q).
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Nat Commun
Hypoxia-induced conversion of sensory Schwann cells into repair cells is regulated by HDAC8. [Abstract]2025 Jan 9;16(1):515. PMID: 39779705 -
Nat Commun
2020 Jan 3;11(1):71. PMID: 31900415 -
Cell Death Differ
Membrane-bound TNF mediates microtubule-targeting chemotherapeutics-induced cancer cytolysis via juxtacrine inter-cancer-cell death signaling. [Abstract]2020 May;27(5):1569-1587. PMID: 31645676 -
Adv Sci (Weinh)
2025 Aug 4:e03972. PMID: 40755418 -
J Clin Invest
Symbiotic exclusivity between CLOCK and TFPI2 drives stemness and immunosuppression in glioblastoma models. [Abstract]2026 Mar 17:e199056. PMID: 41842961 -
J Clin Invest
S1PR1 inhibition induces pro-apoptotic signaling in T cells and limits humoral responses within lymph nodes. [Abstract]2024 Jan 9;134(4):e174984. PMID: 38194271 -
J Exp Clin Cancer Res
Matrix stiffness-upregulated LOXL2 promotes fibronectin production, MMP9 and CXCL12 expression and BMDCs recruitment to assist pre-metastatic niche formation. [Abstract]2018 May 4;37(1):99. PMID: 29728125
JNK-IN-8 purchased from MedChemExpress. Usage Cited in: J Exp Clin Cancer Res. 2018 May 4;37(1):99. [Abstract]
A co-inhibitor of JNK and integrin β obviously decreases phosphorylation levels of JNK and c-JUN and expression of LOXL2 as compared with JNK inhibitor alone or integrin β1 alone.
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Cell Discov
ASCT2 palmitoylation regulated by JNK1-ZDHHC14 axis orchestrates glutamine metabolism and NSCLC progression. [Abstract]2026 Feb 24;12(1):13. PMID: 41730846 -
Cell Rep Med
HE4 drives PD-L1 expression in myeloid cells via IFN-γR-JAK-STAT3 signaling to promote tumor immune evasion. [Abstract]2026 Apr 21;7(4):102691. PMID: 41861828 -
Cell Rep Med
scRNA-seq reveals an immune microenvironment and JUN-mediated NK cell exhaustion in relapsed T-ALL. [Abstract]2025 Apr 25:102098. PMID: 40306275 -
J Neuroinflammation
STAT3 S727 phosphorylation drives pathogenic Th17 differentiation and neuroinflammation in autoimmune disease. [Abstract]2025 Oct 28;22(1):246. PMID: 41152886
JNK-IN-8 purchased from MedChemExpress. Usage Cited in: J Neuroinflammation. 2025 Oct 28;22(1):246. [Abstract]
(K–N) Flow cytometry analysis of IFN-γ and T-bet expression in CD4+ T cells cultured under pTh17 conditions with or without JNK inhibitor (JNK-IN-8) ( 1 μM; 4 days).
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Acta Biomater
Engineered exosome-like nanovesicles suppress tumor growth by reprogramming tumor microenvironment and promoting tumor ferroptosis. [Abstract]2021 Nov;135:567-581. PMID: 34506976 -
Proc Natl Acad Sci U S A
ALPK1 mutants causing ROSAH syndrome or Spiradenoma are activated by human nucleotide sugars. [Abstract]2023 Dec 12;120(50):e2313148120. PMID: 38060563 -
Dev Cell
Endoderm development requires centrioles to restrain p53-mediated apoptosis in the absence of ERK activity. [Abstract]2021 Dec 20;56(24):3334-3348.e6. PMID: 34932949 -
Cell Syst
Systematic analysis of the MAPK signaling network reveals MAP3K-driven control of cell fate. [Abstract]2022 Oct 30;S2405-4712(22)00430-6. PMID: 36356576 -
J Transl Med
TGF-β/JNK axis mediates mitochondrial damage and macrophage cGAS-STING activation in liver Mallory-Denk body pathogenesis. [Abstract]2025 Jun 3;23(1):621. PMID: 40462149
JNK-IN-8 purchased from MedChemExpress. Usage Cited in: J Transl Med. 2025 Jun 3;23(1):621. [Abstract]
IF demonstrates K8 and p62 protein changes after DMSO and JNK-IN-8 ( 3 μM; 24 h) treatment based on JNK pathway activation. Image J software was used to calculate fuorescence intensity of K8 and p62 in each group, and the co-localization of K8 and p62 was expressed as the ratio of the fuorescence intensity of K8 to that of p62. Then 3 randomly selected horizons were analyzed for fuorescence intensity and statistics. Bar =100 μm.
JNK-IN-8 purchased from MedChemExpress. Usage Cited in: J Transl Med. 2025 Jun 3;23(1):621. [Abstract]
The damage of cells, lysosomes and mitochondria in the control group, the TNF-α combined with IFN-γ and TGF-β stimulation group (treated with DMSO), and the TNF-α combined with IFN-γ and TGF-β stimulation group (treated with JNK-IN-8(3 μM; 24 h)) was observed by TEM. Bar = 2 μm/500 μm.
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Cell Rep
Proteomics reveal cap-dependent translation inhibitors remodel the translation machinery and translatome. [Abstract]2021 Oct 12;37(2):109806. PMID: 34644561 -
Clin Transl Med
Tumour cells are sensitised to ferroptosis via RB1CC1-mediated transcriptional reprogramming. [Abstract]2022 Feb;12(2):e747. PMID: 35220675 -
Sci Signal
2026 Jan 13;19(920):eadw3709. PMID: 41529098 -
Sci Signal
The kinase ERK plays a conserved dominant role in the heterogeneity of epithelial-mesenchymal transition in pancreatic cancer cells. [Abstract]2025 Aug 12;18(899):eads7002. PMID: 40794842 -
J Ethnopharmacol
Huaier suppresses cisplatin resistance in non-small cell lung cancer by inhibiting the JNK/JUN/IL-8 signaling pathway. [Abstract]2024 Jan 30;319(Pt 2):117270. PMID: 37832810 -
Cells
IL-3-Induced Immediate Expression of c- fos and c- jun Is Modulated by the IKK2-JNK Axis. [Abstract]2022 Apr 25;11(9):1451. PMID: 35563758 -
Life Sci
OASL activates MAPK to drive psoriatic pathogenesis: Astilbin targeting this axis improves metabolic-inflammation crosstalk. [Abstract]2025 May 11:123698. PMID: 40360089 -
Commun Biol
A genome-scale CRISPR Cas9 dropout screen identifies synthetically lethal targets in SRC-3 inhibited cancer cells. [Abstract]2021 Mar 25;4(1):399. PMID: 33767353 -
Cancer Gene Ther
b-AP15 enhances TRAIL-induced cell death in HNSCC via the induction of ROS/JNK/DR5 signalling. [Abstract]2026 May 2. PMID: 42069980 -
Int J Mol Sci
The Possible Connection of Two Dual Function Processes: The Relationship of Ferroptosis and the JNK Pathway. [Abstract]2022 Sep 20;23(19):11004. PMID: 36232313 -
Expert Opin Ther Targets
Arenobufagin induces cell apoptosis by modulating the cell cycle regulator claspin and the JNK pathway in nasopharyngeal carcinoma cells. [Abstract]2024 May;28(5):461-471. PMID: 38659296 -
J Dairy Sci
Sodium butyrate promotes lipopolysaccharide-induced innate immune responses by enhancing mitogen-activated protein kinase activation and histone acetylation in bovine mammary epithelial cells. [Abstract]2020 Dec;103(12):11636-11652. PMID: 33010913 -
Poult Sci
17β-estradiol mediates sex-biased progesterone receptor expression via estrogen receptor α in chicken pituitary. [Abstract]2025 Dec 5;105(1):106210. PMID: 41380332 -
J Cell Mol Med
The Efficacy of Hellebrigenin Against Nasopharyngeal Carcinoma Cells: The Molecular and Bioinformatic Analysis. [Abstract]2025 Jun;29(11):e70624. PMID: 40434251 -
J Cell Mol Med
JUN activation modulates chromatin accessibility to drive TNFα-induced mesenchymal transition in glioblastoma. [Abstract]2022 Aug;26(16):4602-4612. PMID: 35851726 -
Toxicol Sci
Stress-activated pathways mediate PFAS effects on human placental syncytiotrophoblast cells. [Abstract]2026 Jun 5;209(6):kfag060. PMID: 42172632 -
J Biol Chem
Post-translational regulation of mitogen-activated protein kinase phosphatase (MKP)-1 and MKP-2 in macrophages following lipopolysaccharide stimulation: the role of the C termini of the phosphatases in determining their stability. [Abstract]2014 Oct 17;289(42):28753-64. PMID: 25204653
JNK-IN-8 purchased from MedChemExpress. Usage Cited in: J Biol Chem. 2014 Oct 17;289(42):28753-64. [Abstract]
Effect of the JNK inhibitor (JNK IN 8) on the induction of MKP-1 and MKP-2. RAW264.7 cells are first pretreated with vehicle (DMSO), 3 μM JNK IN 8, 10 μM of U0126, or a combination of both JNK IN 8 and U0126, for 30 min, and then stimulated with LPS for 60 min. Representative images are shown in the panels.
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Cell Signal
2023 May:105:110607. PMID: 36690134 -
Mol Med Rep
JNK‑IN‑8 treatment alleviates lipopolysaccharide‑induced acute lung injury via suppression of inflammation and oxidative stress regulated by JNK/NF‑κB signaling. [Abstract]2021 Feb;23(2):150. PMID: 33355369 -
BMC Cancer
Ubiquitin-specific protease-44 inhibits the proliferation and migration of cells via inhibition of JNK pathway in clear cell renal cell carcinoma. [Abstract]2020 Mar 12;20(1):214. PMID: 32164618 -
Toxicol Appl Pharmacol
Cadmium-induced apoptosis through reactive oxygen species-mediated mitochondrial oxidative stress and the JNK signaling pathway in TM3 cells, a model of mouse Leydig cells. [Abstract]2019 Apr 1:368:37-48. PMID: 30796935 -
Oral Dis
2025 Oct 27. PMID: 41146398 -
Mol Cell Biol
The PHLPP1 N-Terminal Extension is a Mitotic Cdk1 Substrate and Controls an Interactome Switch. [Abstract]2021 Feb 23;41(3):e0033320. PMID: 33397691 -
Reprod Sci
E2F8 Transcriptionally Activates DTL to Promote Endometrial Cancer Progression Via the MAPK Pathway. [Abstract]2025 Dec 29. PMID: 41461624 -
Thorac Cancer
GRP75-mediated upregulation of HMGA1 stimulates stage I lung adenocarcinoma progression by activating JNK/c-JUN signaling. [Abstract]2021 May;12(10):1558-1569. PMID: 33755320 -
Leuk Lymphoma
2021 Dec;62(14):3361-3372. PMID: 34355652 -
Appl Bionics Biomech
Inhibition of TNF- α and JNK Signaling Pathway Can Reduce Paclitaxel-Induced Apoptosis of Mouse Cardiomyocytes. [Abstract]2022 Aug 16:2022:8460121. PMID: 36016921 -
Npj Viruses
2025 Sep 18;3(1):69. PMID: 40968155 -
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bioRxiv
ERK plays a conserved dominant role in pancreas cancer cell EMT heterogeneity driven by diverse growth factors and chemotherapies. [Abstract]2025 Feb 9:2025.02.08.637251. PMID: 39975093 -
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Harvard Medical School LINCS LIBRARY
Solvent & Solubility
DMSO : ≥ 100 mg/mL (197.01 mM; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
* "≥" means soluble, but saturation unknown.
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 2 years; -20°C, 1 year. When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 2 years; -20°C, 1 year. When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
Select the appropriate dissolution method based on your experimental animal and administration route.
- For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
- To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for In Vivo experiments, it is recommended to prepare freshly and use it on the same day.
- The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.
Add each solvent one by one: 10% DMSO 40% PEG300 5% Tween-80 45% Saline
Solubility: ≥ 2.5 mg/mL (4.93 mM); Clear solution
This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.
Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
Add each solvent one by one: 10% DMSO 90% (20% SBE-β-CD in Saline)
Solubility: 2.5 mg/mL (4.93 mM); Suspended solution; Need ultrasonic
This protocol yields a suspended solution of 2.5 mg/mL. Suspended solution can be used for oral and intraperitoneal injection.
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
Please enter the basic information of animal experiments:
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-
-
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Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
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%DMSO +
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
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%+
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+%Tween-80 + +
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%Saline +
The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Working solution concentration: 0.22 mg/mL
Method for preparing stock solution: mg drug dissolved in μL DMSO. Stock solution concentration: mg/mL.
1. Take μL DMSO stock solution;
2. Add μL .
μL , mix evenly;
3. Then add μL Tween 80, mix evenly;
4. Then add μL
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
Protocol
A375 cells are pre-treated with 1 μM JNK-IN-8 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, and Protease 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 from ActivX 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].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
HEK-293 cells stably expressing Interleukin Receptor 1 (HEK293-IL1R) are cultured in Dulbecco’s Modified Eagle’s medium (DMEM) supplemented with 10% FBS, 2 mM glutamine and 1×antimycotic/antibiotic solution. Cells are serum starved for 18 h before incubation with DMSO or JNK-IN-8, stimulated with 2 μM Anisomycin for 1h and lysates are clarified by centrifugation for 10 min at 16000 g and 4°C[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
Purity & Documentation
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Data Sheet (275 KB)
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SDS (393 KB)
- English - EN (393 KB)
- Français - FR (393 KB)
- Deutsch - DE (393 KB)
- Norwegian - NO (393 KB)
- Español - ES (393 KB)
- Swedish - SV (393 KB)
- Italian - IT (393 KB)
- Korean - KR (393 KB)
- Portuguese - PT (393 KB)
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Handling Instructions (2659 KB)
References
Complete Stock Solution Preparation Table
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 2 years; -20°C, 1 year. When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.
| Optional Solvent | Concentration Solvent Mass | 1 mg | 5 mg | 10 mg | 25 mg |
|---|---|---|---|---|---|
| DMSO | 1 mM | 1.9701 mL | 9.8505 mL | 19.7009 mL | 49.2523 mL |
| 5 mM | 0.3940 mL | 1.9701 mL | 3.9402 mL | 9.8505 mL | |
| 10 mM | 0.1970 mL | 0.9850 mL | 1.9701 mL | 4.9252 mL | |
| 15 mM | 0.1313 mL | 0.6567 mL | 1.3134 mL | 3.2835 mL | |
| 20 mM | 0.0985 mL | 0.4925 mL | 0.9850 mL | 2.4626 mL | |
| 25 mM | 0.0788 mL | 0.3940 mL | 0.7880 mL | 1.9701 mL | |
| 30 mM | 0.0657 mL | 0.3283 mL | 0.6567 mL | 1.6417 mL | |
| 40 mM | 0.0493 mL | 0.2463 mL | 0.4925 mL | 1.2313 mL | |
| 50 mM | 0.0394 mL | 0.1970 mL | 0.3940 mL | 0.9850 mL | |
| 60 mM | 0.0328 mL | 0.1642 mL | 0.3283 mL | 0.8209 mL | |
| 80 mM | 0.0246 mL | 0.1231 mL | 0.2463 mL | 0.6157 mL | |
| 100 mM | 0.0197 mL | 0.0985 mL | 0.1970 mL | 0.4925 mL |