Immunohistochemistry was performed in formalin-fixed, paraffin-embedded, 4-μm-thick heart sections as described previously [5 (link),7 (link),8 (link),11 (link)]. Myocytes were stained with an anti-α-sarcomeric actin (α-SA) antibody (Millipore-Sigma, St Louis, MO, USA). Myocyte membranes were stained with Rhodamine conjugated wheat germ agglutinin (WGA) (Vector Laboratories, Inc., Burlingame, CA, USA) to facilitate the identification of individual myocytes for analysis of myocyte cross-sectional area and myocyte density by identification of cell borders. Double immunofluorescent staining was performed with specific anti- α-SA and anti-BrdU or anti-IdU (Abcam, Cambridge, MA, USA) antibodies for evaluation of proliferating myocytes. An anti-CD45 (Abcam, Cambridge, MA, USA) antibody was used to identify inflammatory cells. Nuclei were stained with DAPI (4′,6-diamidino-2-phenylindole).
Anti α sa
Manufactured by Abcam
Sourced in United States
Anti-α-SA is a primary antibody that binds to alpha-smooth muscle actin (α-SMA), a cytoskeletal protein found in vascular smooth muscle cells and other cell types. This antibody can be used in various applications, including immunocytochemistry, immunohistochemistry, and Western blotting, to detect and visualize α-SMA in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Rhodamine conjugated wheat germ agglutinin, by Vector Laboratories (1 mentions)
Anti cd45, by Abcam (1 mentions)
Anti integrin β1, by Abcam (1 mentions)
Enhanced chemiluminescence reagent, by Cytiva (1 mentions)
Anti phospho smad2, by Cell Signaling Technology (1 mentions)
Anti smad2, by Cell Signaling Technology (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Zen 2009, by Zeiss (1 mentions)
Anti α mhc, by Abcam (1 mentions)
Anti serca2, by Thermo Fisher Scientific (1 mentions)
Anti ctnt, by Abcam (1 mentions)
3 protocols using anti α sa
1
Immunohistochemical Analysis of Cardiac Tissue
2
Western Blot Analysis of Cell-Scaffold Constructs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blot assays were performed as described previously.18 (link) In brief, the total protein of the cell-scaffold constructs was prepared using lysis buffer (Tiandz, Beijing, People’s Republic of China). For determination of Western blotting by SDS-PAGE, 100 µg total protein was separated by SDS-PAGE electrophoresis, and anti-α-SA (Abcam), anti-β1-integrin (Abcam), anti-phospho-Smad 2 (Cell Signaling Technology, Beverly, MA, USA), and anti-Smad 2 antibodies (Cell Signaling Technology) were used to incubate the polyvinylidene difluoride membrane with total protein. Enhanced chemiluminescence reagent (Amersham, MA, USA) was utilized to detect the labeled proteins. In some experiments, BASCs grown on the scaffolds were incubated in serum-free medium containing anti-β1-integrin antibody (clone Ha2/5; BD Pharmingen, CA, USA) for 1 hour in 37°C, 0.05% CO2 to block signaling mediated by β1-integrin, which was shown by previous studies.22 (link) Then, the treated constructs were cultured in normal medium for 7 days. In some other experiments, constructs were incubated with ALK-specific inhibitor, SB431542 (10 µM), for 7 days. The band intensities were analyzed with the ImageJ software.
3
Immunofluorescence Characterization of Cardiomyocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells fixed with 4% paraformaldehyde and permeabilized with 0.5% Triton X-100 were incubated with anti-cTnT (1:100, Abcam, Cambridge, UK), anti-α-MHC (1:100, Abcam, Cambridge, UK)), anti-α-SA (1:100, Abcam, Cambridge, UK), anti-CACNA1C (1:100, Abcam, Cambridge, UK), and anti-SERCA2 (1:100; Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA), followed by the appropriate secondary antibody conjugated with Alexa Fluor 488 or PE (1:200; Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA), as previously reported [51 (link)]. Nuclei were counterstained with DAPI (Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA). Samples treated only with the secondary antibody were used as controls. Images were acquired using an Axio Vert A1 microscope. Images were analyzed by ZEN 2009 (Carl Zeiss, Oberkochen, Germany).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!