Immunofluorescence staining of tissue sections was performed as described previously 30 (link). Nuclei were counterstained with 4',6-diamidino-2-phenylindole (DAPI; Sigma Aldrich, St. Louis, MO), and 10 randomly selected fields in each sample were imaged by fluorescence microscopy (Olympus, Tokyo, Japan). Anti-cTnT and anti-vimentin antibodies were purchased from Abcam (Cambridge, MA), and fluorescent secondary antibodies were obtained from Servicebio (Wuhan, China).
Fluorescent secondary antibody
Manufactured by Wuhan Servicebio Technology
Sourced in China
Fluorescent secondary antibodies are laboratory reagents used in various immunoassays, such as Western blotting and immunohistochemistry. These antibodies are designed to bind to the primary antibodies that recognize specific target proteins or antigens. The fluorescent labels attached to the secondary antibodies allow for the detection and visualization of the target molecules, enabling researchers to study protein expression, localization, and interactions within biological samples.
Automatically generated - may contain errors
Lab products found in correlation
4 6 diamidino 2 phenylindole (dapi), by Wuhan Servicebio Technology (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Fluorescence microscopy, by Olympus (1 mentions)
Anti α sma ab124964, by Abcam (1 mentions)
Bovine serum albumin (bsa), by Wuhan Servicebio Technology (1 mentions)
Paraformaldehyde (pfa), by Wuhan Servicebio Technology (1 mentions)
Triton x 100, by Wuhan Servicebio Technology (1 mentions)
4 protocols using fluorescent secondary antibody
1
Immunofluorescence Staining of Tissue Sections
2
Histological Analysis of Carotid Artery Injury
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For histological analysis, hematoxylin and eosin (H&E), Elastica van Gieson (EVG), immunofluorescence, and standard immunohistochemical staining were performed, according to the standard protocols; six sections taken from the middle portion of each artery, 28 days after the carotid artery injury, were examined; and the neointimal area, medial area, and neointima/media (NI/M) ratio were calculated. Anti-α-SMA (ab124964) were purchased from Abcam (Cambridge, MA). Anti-Ki67 antibody (GB13030-2) and fluorescent secondary antibodies were obtained from Servicebio (Wuhan, China).
3
Immunofluorescence Staining of Key Proteins
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Col II, Aggrecan, MMP13, iNOS, p65, IκBα, Nrf2 and HO-1 protein levels were detected by immunofluorescence staining. In a nutshell, cells or tissues were immobilised with 4% paraformaldehyde for 15 min and treated with 2% bovine serum albumin for 1 h to reduce background noise. Cells or tissues were labelled with antibody overnight at 4 °C and then incubated with fluorescent secondary antibody (Servicebio, China) for 2 h. Nuclear staining with DAPI (Servicebio, China)15 min. The cytoskeleton was stained with phalloidin (Servicebio, China) 1 h. The samples were viewed uses Confocal Microscope.
4
Immunofluorescence Staining of Cultured Cells
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Cells were seeded in a 24-well plate, followed by removal of the culture medium, rinsing, and fixation with 4% paraformaldehyde (Servicebio). Subsequently, the membrane was made permeable using 0.5% TritonX-100 (Servicebio), blocked with 1% BSA (Servicebio) in PBS, and subsequently exposed to the primary antibody solution as directed, followed by an overnight incubation at 4 °C. Following the washing step, a pre-made fluorescent secondary antibody (Servicebio) was introduced to all wells and left to incubate at room temperature in the absence of light. It is important to mention that DAPI (Servicebio) was applied for nuclear staining, and subsequent visualization and recording of images took place using a fluorescence microscope.
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