Phospho camkii

Manufactured by Thermo Fisher Scientific

Phospho-CaMKII is a laboratory reagent used to detect and measure the phosphorylation state of the Calcium/Calmodulin-Dependent Protein Kinase II (CaMKII) enzyme. It provides a tool for researchers to study the activation and regulation of this important signaling protein.

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Lab products found in correlation

Nf κb p65, by Thermo Fisher Scientific (1 mentions) α actinin, by Thermo Fisher Scientific (1 mentions) Cox 4, by Cell Signaling Technology (1 mentions) Vdac1, by Abcam (1 mentions) α actinin, by Merck Group (1 mentions) Nupage bis tris gel, by Thermo Fisher Scientific (1 mentions) Turboblotter, by Bio-Rad (1 mentions) Flag tag (flag), by Rockland Immunochemicals (1 mentions) Aclys, by Cell Signaling Technology (1 mentions) Camkii, by Abcam (1 mentions) Odyssey fc imager, by LI COR (1 mentions) Mini protean gel, by Bio-Rad (1 mentions) Gapdh, by Cell Signaling Technology (1 mentions) Image studio software, by LI COR (1 mentions) α tubulin, by Merck Group (1 mentions) Gapdh, by Santa Cruz Biotechnology (1 mentions)

3 protocols using phospho camkii

Protein Expression Analysis in Cardiac Tissue

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Ventricular tissue was homogenized in RIPA buffer (10 mmol/L Tris-Cl (pH
8.0), 1mmol/L EDTA, 0.5 EGTA, 1% Triton X-100, 0.1% sodium
deoxycholate, 0.1% SDS, 140mmol/L NaCl) that was supplemented with
various inhibitors: sodium vanadate, leupeptin, aprotinin,
p-nitrophenyl phosphate, and phenylmethylsulfonyl fluoride.
Western blot analysis was performed according to protocols described previously
[9 (link)]. The antibodies
for immunoblotting were as follows: CaMKIIδ (D.M. Bers, UC Davis); GAPDH
(CST); phospho-IKKα/β at its autophosphorylation site
(Ser176/180) (CST); phospho-CaMKII at its autophosphorylation site (Thr286)
(Thermo); NF-κB p65 (CST); α-actinin (CST); RhoGDI (CST); Lamin
A/C (CST); IκBα (CST)

Gray C.B., Suetomi T., Xiang S., Mishra S., Blackwood E.A., Glembotski C.C., Miyamoto S., Westenbrink B.D, & Brown J.H. (2017). CaMKIIδ subtypes differentially regulate infarct formation following ex vivo myocardial ischemia/reperfusion through NF-κB and TNF-α. Journal of molecular and cellular cardiology, 103, 48-55.

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Cardiac Protein Analysis Protocol

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Heart lysates were prepared in RIPA buffer containing protease and phosphatase inhibitors. Heart lysate or cytoplasm and mitochondria fractions were loaded into 4–12% Bis-Tris NuPAGE gels (ThermoFisher) or 4–15% Mini Protean gels (Bio-Rad). Proteins were transferred to nitrocellulose membrane using Turbo-blotter (Bio-Rad). Primary antibodies [CaMKII (Abcam), 1:1000; phospho-CaMKII (Thermo Scientific), 1:1000; VDAC1 (Abcam), 1:2000; FLAG (Rockland), 1:2000; GAPDH (Cell Signaling), 1:10,000; CKmito (Sigma), 1:6000; CoxIV (Cell Signaling), 1:1000; CK-M (Sigma), 1:10,000; α-actinin (Sigma), 1:1000; OxPhos (Abcam), 1:500; AcLys (Cell Signaling), 1:1000; HA (Sigma), 1:5000; SERCA2a (Badrilla), 1:5000; PDH (Abcam), 1:1000] were incubated with the membrane overnight at 4 °C. Secondary antibodies were incubated with the membrane at room temperature for 1 h. Blots were imaged using an Odyssey Fc Imager (Licor). Bands were quantified using Image Studio Software (Licor), coomassie-stained membranes were quantified using Image J software.

Luczak E.D., Wu Y., Granger J.M., Joiner M.L., Wilson N.R., Gupta A., Umapathi P., Murphy K.R., Reyes Gaido O.E., Sabet A., Corradini E., Tseng W.W., Wang Y., Heck A.J., Wei A.C., Weiss R.G, & Anderson M.E. (2020). Mitochondrial CaMKII causes adverse metabolic reprogramming and dilated cardiomyopathy. Nature Communications, 11, 4416.

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Quantitative Western Blot Analysis

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Western blotting was done using monoclonal anti phospho-CaMKII (Thermo Scientific, MA1-047) α-tubulin (Sigma, T9026) and GAPDH (Santa Cruz Biotechnology, FL-335) primary antibodies, followed by secondary anti-mouse antibodies ECL detection. Western blots were analysed by using ImageJ software.

Sanchez-Alonso J.L., Bhargava A., O’Hara T., Glukhov A.V., Schobesberger S., Bhogal N., Sikkel M.B., Mansfield C., Korchev Y.E., Lyon A.R., Punjabi P.P., Nikolaev V.O., Trayanova N.A, & Gorelik J. (2016). Microdomain-Specific Modulation of L-Type Calcium Channels Leads to Triggered Ventricular Arrhythmia in Heart Failure. Circulation Research, 119(8), 944-955.

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