The biopsy specimen of popliteal artery obtained from an APS patient undergoing femoral-popliteal bypass previously reported(17 (link)) was examined by immunofluorescence for deposits of β2-GPI and MBL. The tissue was stained for β2-GPI with biotin-labeled recombinant antibody CH2-deleted MBB2(17 (link)) followed by FITC-labeled streptavidin (Sigma-Aldrich) and for MBL with primary rabbit IgG anti MBL (Sigma-Aldrich) revealed by sheep anti-rabbit CY3-labeled F(Ab’)2 (Sigma-Aldrich). The sections were examined under a Leica DM2000 fluorescence microscope (Leica) equipped with a digital camera (DFC 490; Leica). Images were acquired by using Leica Application Suite Software. Original magnification 200x
Fitc labeled streptavidin
Manufactured by Merck Group
Sourced in United States
FITC-labeled streptavidin is a fluorescently-labeled protein used in various research applications. Streptavidin has a high affinity for the vitamin biotin, and the fluorescein isothiocyanate (FITC) label allows for the detection of biotin-labeled molecules. This product can be utilized in techniques such as flow cytometry, immunohistochemistry, and Western blotting.
Automatically generated - may contain errors
Lab products found in correlation
Application suite software, by Leica (2 mentions)
Dm2000 fluorescence microscope, by Leica (1 mentions)
Dfc490, by Leica (1 mentions)
Fitc labeled rabbit anti goat igg, by Agilent Technologies (1 mentions)
Benzophenone, by Merck Group (1 mentions)
Biotinylated rabbit anti streptavidin, by Rockland Immunochemicals (1 mentions)
Phosphate buffered saline ph 7, by Thermo Fisher Scientific (1 mentions)
2 aminoethyl methacrylate, by Merck Group (1 mentions)
Sylgard 184, by Dow (1 mentions)
Ez link sulfo nhs lc biotin, by Thermo Fisher Scientific (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Abcam (1 mentions)
4 protocols using fitc labeled streptavidin
1
Immunofluorescent Analysis of Arterial Deposits
2
Mesenteric Tissue Analysis for β2GPI Deposits
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The mesenteric tissue was collected from rats at the end of the in vivo experiment.26 (link) Deposits of β2GPI were analyzed using the biotinylated monoclonal antibody MBB2 and FITC-labeled streptavidin (Sigma-Aldrich).27 (link) IgG and C3 were detected using FITC-labeled goat anti-human IgG (Sigma-Aldrich) and goat anti-rat C3 (Cappel/MP Biomedicals) followed by FITC-labeled rabbit anti-goat IgG (Dako), respectively. The slides were examined using a DM2000 fluorescence microscope equipped with a DFC 490 photo camera and Application Suite software (Leica).
3
UV-Grafted PDMS for CD63 Detection
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A PDMS microchip was used as a substrate for UV grafting (Sylgard 184, Dow Corning, Midland, MI, USA). Benzophenone, 2-aminoethyl methacrylate (AEMA), monoclonal mouse anti-CD63 antibody, and FITC-labeled streptavidin were purchased from Sigma-Aldrich (St. Louis, MO, USA). Biotinylated mouse anti-CD63 antibody was purchased from BioLegend (San Diego, CA, USA). Biotinylated rabbit anti-streptavidin (Rockland Immunochemicals, Gilbertsville, PA, USA) and phosphate-buffered saline (PBS, pH 7.4) were purchased from Thermo Fisher Scientific (Waltham, MA, USA). 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) was purchased from FUJIFILM Wako Pure Chemicals Corporation (Osaka, Japan). All other reagents were of analytical grade or higher.
4
Assessing Testicular Blood-Testis Barrier Integrity
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The integrity of the BTB was assessed by using a biotin tracer, as described previously [61 (link)]. Briefly, 100 μL of EZ-LinkTM Sulfo-NHS-LC-Biotin (10 mg/mL in PBS containing 1 mM CaCl2; Cat# 21335, Thermo Fisher Scientific, Waltham, MA, USA) was injected into the testis interstitium. After 30 min, the testes were cut into 10 μm cryosections and fixed in 4% paraformaldehyde for 15 min. The sections were blocked with 5% fetal bovine serum for 1 h and then incubated with FITC-labeled streptavidin (Cat# S3762, Sigma-Aldrich, St. Louis, MO, USA) for 30 min at room temperature. Finally, the sections were added to the mounting medium with DAPI (Cat# ab104139, Abcam, Cambridge, MA, USA) and visualized by fluorescence microscopy (Olympus BX53, Tokyo, Japan).
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