Single cell-dissociated CM were seeded on Synthemax II-coated glass bottom Petri dishes for 2–3 days. Cells were then fixed in 4% paraformaldehyde for 10 min and permeabilized using 0.1% triton X-100 for 30 min followed by blocking in 5% normal goat serum for 60 min at room temperature. After permeabilization and blocking, the cells were incubated overnight at 4°C with primary antibodies, mouse anti-α-Actinin (Sigma), mouse anti-cTNT (Abcam), rabbit anti-cTNT (Abcam), or rabbit anti-cTNI (Abcam), at 1:100. The samples were rinsed 3× with PBST for 30 min and incubated with secondary antibodies (Cy3 anti-rabbit or FITC anti-mouse, Millipore; 1:500) overnight at 4 C, rinsed, applied a drop of Prolong Gold Anti-fade reagent containing DAPI (Life Technology), and covered with a coverslip. The images were acquired with multiphoton laser scanning confocal microscope (Zeiss LSM 510).
Mouse anti ctnt
Manufactured by Abcam
Sourced in Canada
Mouse anti-cTNT is an antibody that specifically recognizes cardiac troponin T (cTNT), a protein found in cardiac muscle cells. It can be used to detect and quantify cTNT in various laboratory applications.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 510, by Zeiss (2 mentions)
Mouse anti α actinin, by Merck Group (1 mentions)
Cy 3 anti rabbit, by Merck Group (1 mentions)
Rabbit anti ctnt, by Abcam (1 mentions)
Prolong gold antifade reagent, by Thermo Fisher Scientific (1 mentions)
Alexa secondary antibodies, by Thermo Fisher Scientific (1 mentions)
2 protocols using mouse anti ctnt
1
Immunocytochemical Characterization of Cardiomyocytes
2
Immunofluorescence Analysis of iPSC-CMs
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Immunofluorescence staining of iPSC-CMs was carried out on day 30 of maturation. The cells were dissociated from the monolayers using STEMdiff™ Cardiomyocyte Dissociation kits (StemCell Technologies) and were plated on Matrigel-treated 13-mm TC Coverslips (Sarstedt, QC, Canada). The cells were fixed in a mixture of 4% paraformaldehyde and 4% sucrose in PBS for 10 min. Then, they were washed and were permeabilized for 30 min at room temperature in a mixture of 0.1% Triton X-100, 1% BSA, and 5% goat serum in PBS. The cells were incubated overnight at 4 °C with the following antibodies: rabbit anti-MLC2v (1/300, Cat #ab89594, Abcam, ON, Canada) and mouse anti-cTnT (1/300, Cat #ab10214, Abcam). The cells were washed and were incubated for 1 h at room temperature with the appropriate Alexa secondary antibodies (1/250, Cat #A11005 and Cat #A11008, ThermoFisher Scientific, USA). DAPI was used to counterstain the nuclei. The cells were observed using a Zeiss LSM confocal microscope.
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