Cardiac tissues were fixed with 4% paraformaldehyde (Nacalai Tesque). Experiments were performed on paraffin sections after antigen retrieval with citrate buffer (pH 6.0) at 121°C for 20 min. The tissues were incubated with primary antibodies including mouse or rabbit anti-cTnT (1:200 dilution; Abcam, Cambridge, UK), rabbit anti-vimentin (1:200, Abcam), mouse anti-sarcomeric α-actinin (1:400; Sigma-Aldrich), rabbit anti-connexin43 (1:100; Abcam), mouse anti-MLC2a (1:100; Synaptic Systems, Goettingen, Germany), rabbit anti-MLC2v (1:200; Proteintech, Rosemont, IL, USA), mouse anti-FN (1:200; Abcam), rabbit anti-laminin (1:30; Sigma-Aldrich), and rabbit anti-collagen type I (1:500; Abcam). The endothelial cells were immunostained with mouse anti-CD31 antibody (1:200; Dako, Glostrup, Denmark). The tissues were permeabilized with 0.2% Triton X (Sigma-Aldrich), and then non-specific reactivity was blocked with 1% BSA. The tissues were labeled by primary antibodies. Secondary antibodies (1:200) such as Alexa Fluor 488- or Alexa Fluor 546-conjugated goat anti-mouse or anti-rabbit IgG (H+L) (Thermo Fisher Scientific) were added to the tissues. Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI) (Life Technologies) and observed by confocal laser scanning microscopy (FLUOVIEW FV10i; Olympus, Tokyo, Japan).
Rabbit anti connexin43
Manufactured by Abcam
Sourced in Germany, United States
Rabbit anti-connexin43 is a primary antibody that specifically targets the connexin43 protein. Connexin43 is a gap junction protein that plays a crucial role in cell-to-cell communication. This antibody can be used in various research applications to detect and study the expression and localization of connexin43 in different cell types and tissues.
Automatically generated - may contain errors
Lab products found in correlation
Fluoview fv10i, by Olympus (1 mentions)
Mouse anti α sarcomeric actinin, by Merck Group (1 mentions)
Rabbit anti collagen type 1, by Abcam (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Paraformaldehyde (pfa), by Nacalai Tesque (1 mentions)
Rabbit anti laminin, by Merck Group (1 mentions)
Rabbit anti vimentin, by Abcam (1 mentions)
Alexa fluor 546 conjugated goat anti mouse, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (1 mentions)
Anti ctnt, by Abcam (1 mentions)
Rabbit anti mlc2v, by Proteintech (1 mentions)
Goat anti mouse igg1 alexa 488, by Thermo Fisher Scientific (1 mentions)
Perm wash, by BD (1 mentions)
Donkey anti rabbit igg alexa 488, by Thermo Fisher Scientific (1 mentions)
Rabbit anti n cadherin, by Abcam (1 mentions)
Mouse anti α actinin, by Abcam (1 mentions)
Anti rabbit igg alexa594, by Thermo Fisher Scientific (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Dapi fluoromount g, by Southern Biotech (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
3 protocols using rabbit anti connexin43
1
Cardiac Tissue Immunofluorescence Imaging
2
Immunofluorescent Staining of Cardiomyocytes
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In preparation for immunofluorescent staining, plated cardiomyocytes were washed once with PBS. The cells were fixed and permeabilized by applying Cytofix/Cytoperm für 20 minutes. After fixation, the cells were washed with Permwash (BD Biosciences) and stained with primary antibody dilutions for rabbit anti-cardiac troponin T (Abcam, 1:200) mouse anti-α-actinin (Abcam, 1:1000), rabbit anti-myosin light chain 2 (MLC2v) (Proteintech, 1:200), rabbit anti-connexin 43 (Abcam, 1:1000) and rabbit anti-N cadherin (Abcam, 1:200) overnight. The next day, the cells were washed and stained with the respective secondary antibodies anti-rabbit IgG Alexa594, goat anti-mouse IgG1 Alexa488, donkey anti-rabbit IgG Alexa488 or goat anti-mouse IgG1 Alexa594 (all Life Technologies, 1:1000). Next, the stained cardiomyocytes were washed, stained with DAPI to visualize nuclei and evaluated with a Zeiss Observer Z1 microscope.
3
Cardiomyocyte Seeding and Characterization on Magnetic Hydrogels
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Cardiomyocytes were seeded on patterned Mgels (with MNP concentrations of 0, 15%, 30%, and 50%) at 2.5 × 105 cells per well in a 12‐well plate. After 5 culture days, cells were fixed in 4% paraformaldehyde solution (Thermo fisher, USA), washed with PBS, and permeabilized with 0.1% triton X‐100 (Sigma‐Aldrich, USA) in ddH2O. Next, samples were blocked with 10% normal goat serum (Jackson Laboratory) in PBS containing 0.1% bovine serum albumin (BSA; Sigma‐Aldrich). Samples were then incubated overnight at room temperature in a humidified chamber with either mouse anti α‐actinin (Sigma‐Aldrich, USA) at 1:200 in PBS+0.1% BSA or rabbit anti connexin 43 (Abcam) at 1:400 in PBS+0.1% BSA. Alexa goat anti‐mouse 594 and goat anti‐rabbit 488 (Thermo fisher, USA), diluted at 1:500, were used as secondary antibodies and incubated for 1 h. Samples were washed with PBS and mounted with DAPI Fluoromount G (SouthernBiotech, USA). Image acquisition was performed on an Olympus FV3000 confocal microscope (Olympus, Japan), with a UPLXAPO ×60 objective/1.42 NA, equipped with FLUOVIEW acquisition and analysis software.
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