Fluorescence conjugated secondary antibody

Manufactured by Beyotime

Sourced in China

Fluorescence-conjugated secondary antibody is a laboratory reagent used to detect and visualize target proteins or molecules in various biological assays. It consists of a secondary antibody that is chemically linked to a fluorescent dye, enabling the detection and localization of the target of interest through fluorescence microscopy or other fluorescence-based techniques.

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Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Keygen Biotech (9 mentions) Eu5888, by Leica (8 mentions) Fluorescence microscope, by Leica (5 mentions) Col1a1, by Leica (4 mentions) Anti runx2, by Cell Signaling Technology (4 mentions) 4 6 diamidino 2 phenylindole (dapi), by Beyotime (3 mentions) Fluorescence microscopy, by Leica (3 mentions) Col1a1, by Abcam (3 mentions) E cadherin, by Abcam (2 mentions) N cadherin, by Abcam (2 mentions) Eclipse c1, by Nikon (2 mentions) Anti e cadherin, by Abcam (2 mentions) Anti vimentin, by Abcam (2 mentions) Anti cramp, by Abcam (2 mentions) Paraformaldehyde (pfa), by Merck Group (2 mentions) Anti runx2, by Abcam (2 mentions) P erk, by Leica (2 mentions) β catenin, by Cell Signaling Technology (2 mentions) Anti p erk, by Cell Signaling Technology (2 mentions) Anti col1a1, by Abcam (2 mentions)

24 protocols using fluorescence conjugated secondary antibody

NEAT1 Regulation of Tumor Growth

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Female BALB/c nude mice 4–5 weeks of age were purchased from the Shanghai Laboratory Animal Center at the Chinese Academy of Sciences. All experiments were performed in accordance with relevant institutional and national guidelines and the regulations of the Shanghai Medical Experimental Animal Care Commission. Mice (5 per group) were injected subcutaneously with 0.2 mL of a cell suspension containing 5 × 10⁵ cells (the piLenti-shRNA-VECTOR and piLenti-shRNA-NEAT1 stable A2780 cell lines) in the right axilla. The tumor growth rates were monitored. When a tumor was palpable, it was measured every other day, and its volume was calculated according to the formula: volume = length × width² × 0.5.
To investigate the relationship between NEAT1 and LIN28B in vivo, frozen sections from animal experimental tumors were washed with PBS and fixed with 4% paraformaldehyde for 20 min at room temperature. After three washes with PBS, the sections were blocked in 5% goat serum for 1 h. The cells were subsequently incubated with primary antibodies specific for Ki-67 (1:100, Proteintech) and LIN28B (1:50, Abcam; ab71415) overnight at 4 °C. The next day, the sections were washed with PBS and then incubated with fluorescence-conjugated secondary antibodies (Beyotime, China), followed by DAPI. Images were captured under a fluorescence microscope.

Yong W., Yu D., Jun Z., Yachen D., Weiwei W., Midie X., Xingzhu J, & Xiaohua W. (2018). Long noncoding RNA NEAT1, regulated by LIN28B, promotes cell proliferation and migration through sponging miR-506 in high-grade serous ovarian cancer. Cell Death & Disease, 9(9), 861.

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Immunofluorescence Analysis of Cardiomyocytes

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For immunofluorescence analysis of NRCMs, cultured NRCMs were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.1% Triton X‐100 in PBS for 10 minutes, blocked with 3% BSA solution for 1 hour and incubated with an anti‐actin (α‐sarcomeric) antibody (Sigma‐Aldrich; 1:200) or a microtubule‐associated protein 1 light chain 3 (LC3) A/B antibody (Cell Signaling Technology; 1:200) overnight at 4°C. Then, the cells were washed and stained with fluorescence‐conjugated secondary antibodies (Beyotime Biotechnology; Alexa Fluor 647 or Alexa Fluor 488, respectively). Immunofluorescence was analysed with a Nikon A1 confocal microscope (Nikon Corporation), and the cell surface areas and numbers of LC3 puncta were measured using Image‐Pro Plus 6.0 software.

Wang B., Shen D., Tang J., Li J., Xiao Y., Chen X., Cao C., Han D., Gao E., Zhao W., Zhang J, & Chang J. (2019). Sodium (±)‐5‐bromo‐2‐(α‐hydroxypentyl) benzoate ameliorates pressure overload‐induced cardiac hypertrophy and dysfunction through inhibiting autophagy. Journal of Cellular and Molecular Medicine, 23(9), 6048-6059.

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Immunofluorescence Staining of Cell Markers

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For Immunofluorescence staining, cells were grown onto coverslips and fixed in 4% paraformaldehyde for 10 min, and blocked with in 0.05% Triton X-100 and 3% bovine serum albumin (BSA) for 30 min. The coverslips were incubated primary antibodies (N-cadherin (1:100; Abcam, Cambridge, United Kingdom), E-cadherin (1:100; Abcam, Cambridge, United Kingdom), acetylated smad4 (1:100; Proteintech, Wuhan, China)) overnight at 4 °C, followed by fluorescence-conjugated secondary antibodies (Beyotime) for 2 h at room temperature, and nuclei were stained with DAPI (Beyotime) for 5 min.

Li W., Zhu D, & Qin S. (2018). SIRT7 suppresses the epithelial-to-mesenchymal transition in oral squamous cell carcinoma metastasis by promoting SMAD4 deacetylation. Journal of Experimental & Clinical Cancer Research : CR, 37, 148.

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Immunofluorescence Characterization of Cell Adhesion and Proliferation

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EC109 and EC9706 cells were seeded onto sterile coverslips (WHB‐24‐CS, diameter: 14 mm) at a density of 5 × 10⁵ cells/coverslip and incubated for 4 h. Cells were then fixed with 4% formaldehyde and coverslips with fixed cells incubated overnight at 4°C with antibodies specific to E‐cadherin (1:100; Abcam), N‐cadherin (1:100; Abcam), Ki‐67 (1:100; Abcam) and MMP10 (1:100; Abcam). Fluorescence‐conjugated secondary antibodies (1:100; Beyotime) were used for 2 h at room temperature. DAPI (Beyotime, China) was used to stain the nuclei.³, ²⁹

Kong D., Long D., Liu B., Pei D., Cao N., Zhang G., Xia Z, & Luo M. (2021). Downregulation of long non‐coding RNA LOC101928477 correlates with tumor progression by regulating the epithelial‐mesenchymal transition in esophageal squamous cell carcinoma. Thoracic Cancer, 12(9), 1303-1311.

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Immunofluorescent Staining of H9C2 Cells

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H₉C₂ cells were plated in 35 mm dishes, washed with PBS and then fixed with 4% paraformaldehyde for 20 mins at room temperature. After being washed thrice with PBS, the H₉C₂ cells were blocked in 5% goat serum for 1 h. The cells were subsequently incubated with primary antibodies specific for Col1A1 (1:400, Abcam) overnight at 4°C. On the next day, the H₉C₂ cells were washed with PBS and incubated with fluorescence-conjugated secondary antibodies (Beyotime, China) followed by DAPI. Images were captured under a fluorescence microscope.

Gong H., Lyu X., Dong L., Tan S., Li S., Peng J., Liu Y, & Zhang X. (2022). Obstructive Sleep Apnea Impacts Cardiac Function in Dilated Cardiomyopathy Patients Through Circulating Exosomes. Frontiers in Cardiovascular Medicine, 9, 699764.

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Regulation of Epithelial-Mesenchymal Transition

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NCI-H292 cells were seeded on chamber slides in serum-free medium. Once the cells reached near 80% confluence, they were incubated with CSE (5%) or LL-37 synthetic peptide (5 µg/mL) with or without pretreatment with EGFR inhibitor AG1478, TGF-α neutralized antibodies, and TACE inhibitor TAPI-1. The slides were then washed in PBS and fixed in 4% paraformaldehyde for 20 min. Next, the cells were permeabilized with Triton X100 (1%) for 20 min and further incubated with corresponding primary antibodies overnight. The following antibodies were used at appropriate dilutions: anti-CRAMP (1:50), anti-E-cadherin (1:100), and anti-vimentin (1:100) (Abcam, Cambridge, USA). The slides were incubated with fluorescence-conjugated secondary antibodies (Beyotime, Shanghai, China) the next day for 1 h at room temperature. Images were taken using fluorescence microscopy (ECLIPSE C1, Nikon, Japan) with 200× magnification.

Jiang Z., Zhang Y., Zhu Y., Li C., Zhou L., Li X., Zhang F., Qiu X, & Qu Y. (2021). Cathelicidin induces epithelial-mesenchymal transition to promote airway remodeling in smoking-related chronic obstructive pulmonary disease. Annals of Translational Medicine, 9(3), 223.

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Immunofluorescence Staining of SIRT-1 and PARP-1

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MCs on 35 mm glass dishes were treated as described previously.50 (link) The plates were washed with cold PBS and fixed with 4% paraformaldehyde for 20 min at room temperature. After permeabilizing with 0.3% of triton, the unspecific sites of the cells were blocked by 1% BSA + 0.3% triton. Then, the cells were incubated with primary antibody experimenters used before,35 (link) rabbit anti-SIRT-1 diluted 1:10 (Santa Cruz Biotechnology) and mouse anti-PARP-1 diluted 1:10 (Enzo Life Sciences, Farmingdale NY) overnight at 4°C and incubated with fluorescence conjugated secondary antibodies (Beyotime, China) counterstained with DAPI next day. Images were analyzed under a fluorescence-inverted microscope.

Zhu H., Fang Z., Chen J., Yang Y., Gan J., Luo L, & Zhan X. (2021). PARP-1 and SIRT-1 are Interacted in Diabetic Nephropathy by Activating AMPK/PGC-1α Signaling Pathway. Diabetes, Metabolic Syndrome and Obesity: Targets and Therapy, 14, 355-366.

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Airway Remodeling Assessment in Mice

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The lung tissue collected from the mice experimental groups was fixed, cut into 4-µm sections, and stained with Masson’s trichrome staining to assess airway remodeling. Immunofluorescence assays were conducted as described in previous publications (22 (link)). Briefly, the slides were blocked with 5% bovine serum albumin (Sigma, Darmstadt, Germany) for 1 h at room temperature and then incubated with primary antibodies at 4 °C overnight. The following primary antibodies were used at the appropriate dilutions: anti-CRAMP (1:50), anti-E-cadherin (1:100), and anti-vimentin (1:100) (Abcam, Cambridge, USA). The slides were incubated the next day with fluorescence-conjugated secondary antibodies (Beyotime, Shanghai, China) for 1 h at room temperature. Then, the slides were examined and photographed using fluorescence microscopy (ECLIPSE C1, Nikon, Japan) with 40× magnification. The fluorescence intensities of CRAMP, E-cadherin, and vimentin were calculated using Image-Pro Plus 6.0 software and shown as IOD/area.

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Immunofluorescence Staining of Rat Kidney

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Rat kidneys were harvested, frozen in Tissue-Tek OCT media, and sliced sequentially into 10 μm sections using a cryostat. Sections were permeabilized with 0.1% Triton X-100 in phosphate-buffered saline (PBS) for 20 min and then blocked with 5% donkey serum for 1 h and incubated with rabbit anti-E-cadherin (1:100; 20874-1-AP; Proteintech) overnight at 4°C. Next, sections were incubated for 5 min with 4′,6-diamidino-2-phenylindole (DAPI) and then for 2 h with fluorescence-conjugated secondary antibodies (Beyotime, Shanghai, China) in a dark room at room temperature. Slices were visualized under a fluorescence microscope (Leica Microsystems, Germany).

Luo M., Luo S., Xue Y., Chang Q., Yang H., Dong W., Zhang T, & Cao S. (2023). Aerobic exercise inhibits renal EMT by promoting irisin expression in SHR. iScience, 26(2), 105990.

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Immunofluorescence Analysis of Aorta Oxidation

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Aorta sections were permeabilized with 0.1% Triton X-100 in PBS for 20 min, blocked with 5% goat serum for 1 h, and incubated with antibodies overnight at 4°C. The tissue sections were then incubated for 5 minutes with 4′,6-diamidino-2-phenylindole (DAPI) and then for 1 h with fluorescence-conjugated secondary antibodies (Beyotime, Shanghai, China) in the dark at 37°C. Tissues were observed by fluorescence microscopy (Leica Microsystems, Germany). The antibodies used were 8-OHdG (1 : 200, bs-1278R, Bioss), NLRP3 (1 : 200, NBP2-12446, NOVUS), and GSDMD (1 : 200, NBP2-33422, NOVUS). Fluorescence intensity was quantified using the ImageJ 1.8 software.

Luo M., Hu Y., Lv D., Xie L., Yang S., Zuo D., Xue Y, & He A. (2022). Recurrent Hypoglycemia Impaired Vascular Function in Advanced T2DM Rats by Inducing Pyroptosis. Oxidative Medicine and Cellular Longevity, 2022, 7812407.

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