After the paraffin tissue sections were deparaffinized, they were placed in citrate buffer solution, heated 20 min and then cooled to room temperature. After being incubated with endogenous peroxidase blocking solution for 10 min, the sections were washed three times with PBS for 10 min each time, 5% BSA was added for 1 h at room temperature, and the following primary antibodies were added and incubated overnight at 4 °C: smooth muscle actin (1:200, Proteintech, Rosemont, IL), vimentin (1:200, Proteintech, Rosemont, IL), NLRP3 (1:100, Proteintech, Rosemont, IL), caspase 1/P20/P10, (1:200, Proteintech, Rosemont, IL), and IL-1 beta (1:200, Abcam, Cambridge, UK). The sections were washed three times with PBS for 10 min each time, and the reaction-enhancing solution was added. An appropriate amount of peroxidase-labeled goat anti-rabbit IgG was added and incubated at room temperature for 20 min. After being washed three times with PBS for 10 min each time, DAB was added to the sample for several seconds. After observing the color change under the light microscope, the reaction was terminated by placing the sample in deionized water. The nuclei were stained with hematoxylin, and the slides were examined under the light microscope.
Smooth muscle actin
Manufactured by Proteintech
Sourced in United States
Smooth muscle actin is a protein that is a component of the contractile apparatus in smooth muscle cells. It is a member of the actin family and plays a crucial role in the contraction and movement of smooth muscle tissues.
Automatically generated - may contain errors
Lab products found in correlation
Il 1 beta, by Abcam (1 mentions)
Vimentin, by Proteintech (1 mentions)
Caspase 1 p20 p10, by Proteintech (1 mentions)
Nlrp3, by Proteintech (1 mentions)
Hsp70, by Abcam (1 mentions)
Hif 1α, by Proteintech (1 mentions)
E cadherin, by Proteintech (1 mentions)
Enhanced chemi luminescence (ecl), by Abbkine (1 mentions)
Collagen type 1, by Proteintech (1 mentions)
β tubulin, by Cell Signaling Technology (1 mentions)
Ripa buffer, by Beyotime (1 mentions)
Gapdh, by Abcam (1 mentions)
Polyvinylidene difluoride membrane, by Merck Group (1 mentions)
P akt, by Proteintech (1 mentions)
2 protocols using smooth muscle actin
1
Immunohistochemical Analysis of NLRP3 Inflammasome
2
Western Blot Analysis of Protein Markers
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Cell and tissue protein was extracted using RIPA buffer (Beyotime, China). Proteins (20-30 mg) were separated by 10% sodium dodecyl-sulfate polyacrylamide gel electrophoresis gel before being transferred to polyvinylidene difluoride membranes (Merck Millipore, USA). After being blocked in 5% milk for 1 h, the membranes were then probed at 4°C overnight with antibodies against PTEN (1:500; Santa Cruz Biotechnology, USA, sc-7974), AKT (1:1000; Proteintech Group, USA, 10176-2-AP), p-AKT (1:1000; Proteintech Group, USA, 66444-1-lg), HIF-1α (1:1000; Proteintech Group, USA, 20960-1-AP), MMP9 (1:1000; Proteintech Group, USA, 10375-2-AP), Collagen Type I (1:1000; Proteintech Group, USA, 14695-1-AP), E-cadherin (1:1000; Proteintech Group, USA, 20874-1-AP), Smooth Muscle Actin (1:1000; Proteintech Group, USA, 14395-1-AP), CD63 (1:1000; Proteintech Group, USA,25682-1-AP), CD81 (1:1000; Proteintech Group, USA, 66866-1-lg), HSP70 (1:1000; Abcam , USA, ab5439), β-Tubulin (1:5000; Cell Signaling Technology, USA, 2146) and GAPDH (1:5000; Abcam , USA, ab8245). The membranes were washed by TBS with Tween, then incubated with horseradish peroxidase-conjugated secondary antibodies for 1 h. After that, the membranes were immersed in ECL (Abbkine, China) and visualized using a chemiluminescence instrument.
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