The cells were placed in 6-well plates at a density of 1.0×105 cells/ml and 2 ml/well. Following treatment with AngII, the cardio-myocytes were fixed in 4% paraformaldehyde, permeabilized with 0.1% Nonidet P-40, and immunostained with mAKAPβ antibody and α-actinin antibody (Cat. no. sc-17829; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA; in order to distinguish the cardiomyocytes from the contaminating fibroblasts) in PBS [0.1% Nonidet P-40 and 3% bovine serum albumin (BSA)] followed by Alexa fluorescent dye-conjugated specific secondary antibody and Hoechst 33258 DNA staining, as previously described (29 (link)). Fluorescent images were taken using a Leica DMRA fluorescence microscope (Leica Microsystems GmbH, Wetzlar, Germany) and the cell surface areas were calculated using the NIH ImageJ image processing program. Data were pooled from 2 independent experiments.
α actinin antibody
Manufactured by Santa Cruz Biotechnology
Sourced in United States
The α-actinin antibody is an affinity-purified polyclonal antibody that recognizes α-actinin, a cytoskeletal protein involved in the organization of actin filaments. This antibody can be used for the detection and localization of α-actinin in various applications, such as Western blotting, immunohistochemistry, and immunocytochemistry.
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2 protocols using α actinin antibody
1
Immunostaining of Cardiomyocytes with AngII
2
Quantifying Cardiomyocyte Apoptosis in I/R
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The apoptosis of cardiomyocytes in I/R mouse models was analyzed by terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) staining with an In Situ Cell Death Detection Kit (Roche, Basel, Switzerland). Briefly, cardiac sections (5 μm) were fixed in 4% paraformaldehyde, permeabilized with 0.1% Triton X-100, followed by incubation with TUNEL reaction mixture in the dark for 1 h at 37°C. The cell nuclei were counterstained with DAPI (1:100; Molecular Probes, Eugene, Ore). In addition, myocardium was specifically labeled by α-actinin antibody (1:100; Santa Cruz). Images were obtained by using a fluorescence microscope. Apoptotic index was calculated as the number of TUNEL positive nuclei/total number of nuclei × 100% with blinded manners.
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