Ab136975

Manufactured by Abcam

Ab136975 is a polyclonal antibody that recognizes a specific target protein. The antibody is designed for use in various laboratory techniques, such as Western blotting and immunohistochemistry. This product is suitable for research purposes only.

Automatically generated - may contain errors

Lab products found in correlation

Ab179813, by Abcam (1 mentions) Hpa037480, by Merck Group (1 mentions) Tissuelyser lt bead mill, by Qiagen (1 mentions) Ab87399, by Abcam (1 mentions) Ab14746, by Abcam (1 mentions) Hpa016480, by Merck Group (1 mentions) T6199, by Merck Group (1 mentions) Las 4000, by Fujifilm (1 mentions)

2 protocols using ab136975

Mitochondrial and Circadian Protein Analysis

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Heart apex was pulverized in tissue pulverizer cooled in dry ice. Pulverized tissue was placed in 600 μL fresh lysis buffer (150 mM Na₂CO₃, supplemented with 1X protease/phosphatase inhibitor cocktail, pH 11) in a 2 ml Qiagen tube with one metal bead; samples were homogenized for 5 min at 50 oscillations/second in a Qiagen Tissuelyser Lt Bead Mill. Samples were let to settle on ice for 30 min and subsequent sonication (20 sec at 40% sonication intensity). 5 μg protein was loaded per well and resolved in 12% SDS gel. For immunoblot detection of mitochondrial and circadian proteins antibodies against ND1 (SAB4501927, Sigma), MCU (14997, Cell Signaling), MCUb (HPA048776, Sigma), MICU1 (HPA037480, Sigma), MICU2 (ab101564, Abcam), NCLX (ab136975, Abcam), EMRE (PAS23032, custom), BMAL1 (14020, Cell Signaling), Per2 (ab179813, Abcam), and GAPDH (2118S, Cell Signaling) were used at 1:1000 dilution. Band density analysis was performed with ImageJ software; all data is represented as total protein expression normalized by GAPDH.

Abdel-Rahman E.A., Hosseiny S., Aaliya A., Adel M., Yasseen B., Al-Okda A., Radwan Y., Saber S.H., Elkholy N., Elhanafy E., Walker E.E., Zuniga-Hertz J.P., Patel H.H., Griffiths H.R, & Ali S.S. (2021). Sleep/wake calcium dynamics, respiratory function, and ROS production in cardiac mitochondria. Journal of Advanced Research, 31, 35-47.

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Quantitative Protein Expression Analysis

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Protein samples were extracted with non-reducing Laemmli buffer without bromophenol blue and quantified by Bradford assay. Extracts were loaded onto 10% standard polyacrylamide gel electrophoresis after adding 5% 2-mercaptoethanol, and subsequently transferred to nitrocellulose membranes or PVDF membranes. The following antibodies were used: polyclonal anti-NCLX (ab136975, Abcam; ARP44042_P050, Aviva Systems Biology), monoclonal anti-Fp70 (459200, Invitrogen), monoclonal anti-NDUFS4 (ab87399, Abcam), monoclonal anti-RISP (UQCRFS1) (ab14746, Abcam), polyclonal anti-MCU (HPA016480, Sigma-Aldrich) and monoclonal anti-α-tubulin (T6199, Sigma). Antibody binding was detected by chemiluminescence with species-specific secondary antibodies labelled with horseradish peroxidase (HRP), and visualized on a digital luminescent image analyser (Fujifilm LAS-4000), with the exception of ab136975 and 459200, which were detected by fluorescence as previously described^{43 (link)}.

Hernansanz-Agustín P., Choya-Foces C., Carregal-Romero S., Ramos E., Oliva T., Villa-Piña T., Moreno L., Izquierdo-Álvarez A., Cabrera-García J.D., Cortés A., Lechuga-Vieco A.V., Jadiya P., Navarro E., Parada E., Palomino-Antolín A., Tello D., Acín-Pérez R., Rodríguez-Aguilera J.C., Navas P., Cogolludo Á., López-Montero I., Martínez-del-Pozo Á., Egea J., López M.G., Elrod J.W., Ruíz-Cabello J., Bogdanova A., Antonio Enríquez J, & Martínez-Ruiz A. (2020). Na+ controls hypoxic signalling by the mitochondrial respiratory chain. Nature, 586(7828), 287-291.

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