Fluorescently labeled secondary antibodies

Manufactured by Proteintech

Fluorescently labeled secondary antibodies are laboratory reagents designed to detect and visualize target proteins in various experimental techniques, such as Western blotting, immunohistochemistry, and flow cytometry. These antibodies are conjugated with fluorescent dyes, enabling the detection and localization of proteins of interest through fluorescence-based methods.

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Lab products found in correlation

Anti gapdh mouse monoclonal antibody, by Proteintech (1 mentions) Odyssey infrared imaging system, by LI COR (1 mentions) Bca protein assay kit, by GenStar (1 mentions) Fluorescence microscope, by Olympus (1 mentions)

2 protocols using fluorescently labeled secondary antibodies

Lung Tissue Protein Extraction and Quantification

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The lung tissue homogenate was collected then freeze-thawed repeatedly 3–5 times using liquid nitrogen. After being centrifuged at 5000× g for 7 min, the supernatant was protein extracts. Sample protein concentration was determined with the BCA Protein Assay Kit (Genstar). The supernatants were then divided according to the sample protein concentration. Sample loading buffer was added to the supernatants and boiled for 5 min. Samples were then centrifuged, and the supernatant was removed. Approximately 20 μg of sample supernatant was loaded per lane, separated on a precast polyacrylamide Bis-Tris gel with a 4–12% gradient (Solarbio), and transferred onto a nitrocellulose membrane. The membranes were blocked in 10% milk/TBS-T buffer for 1 h at RT and incubated overnight with the following antibodies: rabbit anti-NLRP6 polyclonal antibody (1:2000, Immunoway), rabbit anti-GSDMD polyclonal antibody (1:5000, Proteintech), and mouse anti-GAPDH monoclonal antibody (1:10,000, Proteintech). Membranes were incubated with fluorescently labeled secondary antibodies (1:10,000, Proteintech) at RT for 1 h. The protein bands were detected with the Odyssey^® infrared imaging system (LI-COR Biosciences). The relative expression of each target gene was obtained using the following formula:
relative expression of gene = (density of the gene band)/(density of β-actin band).

Shi B., Han H., Li H., Tan L., Li X., Wang K., Li B., He W., Tian C., Yan F., Shi Y., Zheng Y, & Zhao Z. (2022). NLRP6 Induces Lung Injury and Inflammation Early in Brucella and Influenza Coinfection. Journal of Personalized Medicine, 12(12), 2063.

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Immunofluorescence Analysis of Renal and Cell Samples

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The renal tissues were fixed with 4% paraformaldehyde for 10 min, permeabilized in 0.2% Triton X-100 at room temperature for 10 min, then blocked with goat serum for 30 min at 37 °C. Subsequently, the renal tissues were incubated with antibodies against collagen Ι and TFEB in PBS overnight at 4 °C, and followed by fluorescently labeled secondary antibody for 1 h at 37 °C. After washing five times with PBS, the sections were incubated with DAPI dissolved in PBS for 10 min. Images were captured using a fluorescence microscope (Olympus, Tokyo, Japan).
HK-2 cells were washed twice with PBS and fixed with 4% paraformaldehyde at 4 °C for 10 min. Subsequently, the cells were permeabilized with Triton X-100, sealed with goat serum at room temperature for 30 min, and then incubated with specific primary antibodies against TFEB, LC3B, p62, E-cadherin, and α-SMA at 4 °C overnight, followed by fluorescently labeled secondary antibodies (Proteintech, Chicago, IL) for 1 h at room temperature without light. The cells were then incubated with DAPI for 5 min. Images were obtained using a fluorescence microscope (Olympus, Japan).

Du Y., Wu M., Song S., Bian Y, & Shi Y. (2024). TXNIP deficiency attenuates renal fibrosis by modulating mTORC1/TFEB-mediated autophagy in diabetic kidney disease. Renal Failure, 46(1), 2338933.

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