Kn 62
Manufactured by Cayman Chemical
Sourced in United States
KN-62 is a laboratory compound that functions as a selective and potent inhibitor of the calcium/calmodulin-dependent protein kinase II (CaMKII) enzyme. It is commonly used in research applications to investigate the role of CaMKII in various biological processes.
Automatically generated - may contain errors
Lab products found in correlation
Crocin, by Merck Group (1 mentions)
Kn 92, by Cayman Chemical (1 mentions)
Cobalt protoporphyrin copp, by Enzo Life Sciences (1 mentions)
Actinomycin d actd, by Merck Group (1 mentions)
Cycloheximide (chx), by Merck Group (1 mentions)
Sp600125, by AG Scientific (1 mentions)
Ly294002, by AG Scientific (1 mentions)
Pd98059, by AG Scientific (1 mentions)
Sb203580, by AG Scientific (1 mentions)
Bafilomycin, by Cayman Chemical (1 mentions)
Histamine, by Merck Group (1 mentions)
Fluoromount gtm, by Electron Microscopy Sciences (1 mentions)
Lsm 700 confocal microscope, by Zeiss (1 mentions)
U0126, by LC Laboratories (1 mentions)
Wortmannin, by LC Laboratories (1 mentions)
Kainate, by Bio-Techne (1 mentions)
L glutamate, by Abcam (1 mentions)
Ly294002, by Bio-Techne (1 mentions)
D ap5, by Bio-Techne (1 mentions)
Pd184352, by Merck Group (1 mentions)
5 protocols using kn 62
1
CaMKII Inhibition in NMDA-LTD
2
Investigating Crocin and LPS-Induced Cellular Responses
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Crocin (Figure 1(a) ), LPS (phenol extracted from Salmonella enteritidis), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), actinomycin D (Act. D), cycloheximide (CHX), 2-(3,6-bis(acetyloxy)-2,7-dichloro-9H-xanthen-9-yl)benzoic acid acetoxymethyl ester (Fluo-3/AM), and other reagents were purchased from Sigma-Aldrich (St. Louis, MO, USA). Cobalt protoporphyrin (CoPP) and zinc protoporphyrin IX (ZnPP) were bought from Enzo Life Sciences, Inc. (Farmingdale, NY, USA). LY294002, PD98059, SP600125, and SB203580 were obtained from A.G. Scientific (San Diego, CA, USA). Nifedipine, 1,2-bis (o-aminophenoxy) ethane-N,N,N′,N′-tetraacetic acid tetra (acetoxymethyl) ester (BAPTA/AM), and calmidazolium chloride were acquired from Calbiochem (EMD Millipore, Billerica, MA, USA). KN92, KN93, and KN62 were purchased from Cayman Chemical (Ann Arbor, MI, USA) and Calbiochem.
3
Intracellular pH Measurement in Cell Lines
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Beas-2B, A549, and HEK293T cells were cultured on cover glasses and treated with 100 μM histamine (#H7125, Sigma), 10 μM KN62 (#13318, Cayman, Ann Arbour, MI, USA), and 2 μM bafilomycin (#11038, Cayman) for 24 h. The incubated media were removed and the cells washed with heated a regular solution at 37 °C. pHRodoTM Green-AM as an intracellular pH indicator (#p35373, Invitrogen) was treated to the cells and incubated at 37 °C for 30 min. After removal of the supernatants, fixation was performed with 4% paraformaldehyde at room temperature for 10 min in the dark. The cells were washed with DPBS and the cover glasses were carefully attached and mounted on slide glass with Fluoromount-GTM (Electron Microscopy Sciences) containing 4′,6-diamidino-2-phenylindole (DAPI). To acquire fluorescence images, Zeiss LSM700 confocal microscope (Fluoview, Carl Zeiss, Germany) was used and analysed with ZEN software.
4
Monitoring cAMP-mediated Signaling Pathways
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To monitor the cAMP-mediated signal transduction pathway, CRE-GFP 293 cells in 6-well plates (1 × 106/well) were pretreated with kinase inhibitor H-89 (LC Laboratories, Woburn, MA, USA), KN-62 (Cayman Chemical, Ann Arbor, MI, USA), U0126 (LC Laboratories), or wortmannin (LC Laboratories) (10 μM) for 4 h before forskolin or LM-021 (10 μM) addition for 5 h. GFP fluorescence/protein and total/phosphorylated PKA, CaMKII, ERK, PI3K, and CREB levels were examined. In addition, Aβ-GFP or ∆K280 tauRD-DsRed SH-SY5Y cells were treated with retinoic acid (10 μM) on day 1, followed by the addition of LM-021 (5 or 10 μM) and doxycycline (5 or 2 μg/ml) on day 2, as described. Kinase inhibitors (10 μM) were added on day 6. On day 8, the cells were collected for protein expression analysis of BDNF, BCL2 (BCL2 apoptosis regulator), and BAX (BCL2 associated X, apoptosis regulator), and total/phosphorylated PKA, CaMKII, ERK, PI3K, and CREB. Also, cells were stained with DAPI and assessed for neurite outgrowth as described above.
5
Pharmacological Modulation of Neuronal Signaling
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L-glutamate (Abcam, Cambridge, UK) was dissolved in 1 molar eq. NaOH and used at a final concentration of 1 mM. Excitatory glutamate agonists NMDA (Tocris, Bristol, UK), kainate (Tocris) and AMPA (Abcam) were dissolved in dH 2 O and used at 50 µM. The MEK1/2 inhibitor PD184352 (Sigma, St. Louis, MO, USA), CaMKII inhibitor KN-62 (Cayman Chemical, Michigan, USA), PI3K inhibitor LY294002 (Tocris) and the NMDAR inhibitor D-AP5 (Tocris) were dissolved in DMSO and used at 2 µM, 10 µM, 10 µM and 50 µM, respectively. The cell-permeable phosphopeptide activator of PI3K, 740 Y-P (Tocris), was dissolved in dH 2 O and used at 10 µg/ml. The Na + channel blocker tetrodotoxin (TTX) citrate (Abcam) was dissolved in dH 2 O and used at 2.5 µM. DMSO or NaOH at the corresponding concentrations were used as vehicle for control conditions.
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