Anti fitc alexa fluor 488

Manufactured by Thermo Fisher Scientific

Anti-FITC Alexa Fluor 488 is a fluorescent dye conjugated to an anti-FITC antibody. It is designed for use in flow cytometry, immunofluorescence, and other applications where detection of FITC-labeled targets is required.

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Close spelling variants of this product

FITC-conjugated anti-mouse Alexa Fluor-488 antibody FITC-conjugated anti-mouse Alexa Fluor-488 Goat anti-FITC Alexa Fluor 488 Alexa Fluor 488 (FITC) Alexa Fluor 488 anti Anti-FITC Alexa 488 Alexa Fluor 488 Alexa 488 Alexa Fluors

3 protocols using anti fitc alexa fluor 488

Synovial Cell Immunophenotyping by Flow Cytometry

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The following antibodies and reagents were used for the analysis of synovial cells with flow cytometry and cell sorting: anti-CD45-APC-H7 (2D1, BD Pharmingen), anti-CD235a-APC-Alexa Fluor750 (11E4B-7-6(KC16), Beckman Coulter), anti-CD31-PE-Cyanine7 (WM-59, eBioscience), anti-CD146-BV450 (P1H12, BD Horizon), anti-CD34-PE (4H11, eBioscience), anti-PDPN-PerCP-eFluor710 (NZ-1.3, eBioscience), anti-THY1-FITC (5E10, BD Pharmingen), anti-cadherin-11-biotin (23C6), human TruStain FcX (BioLegend), streptavidin-APC (Jackson ImmunoResearch), Live/Dead fixable aqua dead cell stain kits (Molecular Probes). For immunofluorescence staining of synovial tissue, following antibodies and reagents were used: anti-CD45 (135-4C5, AbD Serotec), anti-CD34 (EP373Y, Abcam), anti-PDPN (NZ-1.3, eBioscience), anti-THY1 (F15-42-1, Merck Millipore, and clone Thy-1A1, R&D Systems), anti-cadherin-11-Biotin (23C6), anti-Ki67 (16A8, BioLegend), anti-mouse IgG1-FITC (Southern Biotech), anti-mouse IgG2a FITC (Southern Biotech), anti-mouse IgG2b-Alexa Fluor 647 (Life Technologies), anti-rat IgG-Alexa Fluor 594 (Life Technologies), anti-rat IgG-Alexa Fluor 647, anti-rabbit IgG-Alexa Fluor 546 (Life Technologies), Hoechst 33258 (Life Technologies), and anti-FITC Alexa Fluor 488 (Life Technologies).

Mizoguchi F., Slowikowski K., Wei K., Marshall J.L., Rao D.A., Chang S.K., Nguyen H.N., Noss E.H., Turner J.D., Earp B.E., Blazar P.E., Wright J., Simmons B.P., Donlin L.T., Kalliolias G.D., Goodman S.M., Bykerk V.P., Ivashkiv L.B., Lederer J.A., Hacohen N., Nigrovic P.A., Filer A., Buckley C.D., Raychaudhuri S, & Brenner M.B. (2018). Functionally distinct disease-associated fibroblast subsets in rheumatoid arthritis. Nature Communications, 9, 789.

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Quantifying Mitochondrial Network Morphology

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Cells were adhered to chamber slides at 2x10³/well and cultured with or without 1ng/ml TNFα for 24 hours. Slides were fixed in acetone, air-dried and stored at -20°C. Slides were rehydrated in PBS, blocked in 10% normal goat serum and incubated with mouse anti-TOMM20 (4F3, Abcam, UK) prior to goat anti-mouse IgG1 (Southern Biotech, Birmingham, USA). The FITC signal was amplified with anti-FITC Alexa Fluor 488 (Life Technologies) and nuclei were stained with Hoechst 33258. Slides were mounted in Prolong Diamond (Life Technologies) before imaging. Images were captured on the Leica DM6000 using the proprietary software and processed using Fiji (34 (link)) in a method adapted from (35 (link)). In brief, the image in the TOMM-20 channel was sharpened, thresholded, converted to a mask and then skeletonized prior to running the binary connectivity plug-in as described (36 (link), 37 (link)). For visualisation the Glasbey lookup table was used and numbers of each pixel connection type were exported. Nuclei were counted as an assessment of cell number in each field of view and these data were combined.

Falconer J., Pucino V., Clayton S.A., Marshall J.L., Raizada S., Adams H., Philp A., Clark A.R., Filer A., Raza K., Young S.P, & Buckley C.D. (2021). Spontaneously Resolving Joint Inflammation Is Characterised by Metabolic Agility of Fibroblast-Like Synoviocytes. Frontiers in Immunology, 12, 725641.

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Immunofluorescence Imaging of Synovial Immune Cells

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6 micron sections of synovium frozen in OCT were fixed in acetone, rehydrated in PBS, and blocked with 10% normal goat serum prior to application of primary antibodies as follows: PD-1 (EH12.2H7, BioLegend), CD3 (SP7, Abcam), CD20 (L26, Dako), CXCR5 (MAB190, R&D Systems), all at a dilution of 1:100 except for CD20, which was used at 1:300. All secondary antibodies were raised in goat. CXCR5 was detected using anti mouse IgG2b biotin (Southern biotech) followed by streptavidin conjugated AlexaFluor 546 (Life Technologies), CD20 with anti-mouse IgG2a FITC (both Southern Biotech), PD-1 with anti-mouse IgG1 conjugated to AlexaFluor 647 and CD3 with anti-rabbit AlexaFluor 546 (both Life Technologies). FITC staining was amplified with anti-FITC AlexaFluor 488 (Life Technologies). Slides were mounted using ProLong Diamond (Life Technologies), left to cure overnight and imaged using a Zeiss LSM 780 confocal microscope. Images were processed using Zen Black (Zeiss) and then ImageJ. Cell counts were performed on images obtained from confocal imaging using the Cell Counter plugin for ImageJ (imagej.net/Cell_Counter). Synovial regions were categorized as ‘lymphoid aggregates’ when the B cells and T cells formed distinct clusters, and ‘diffusely infiltrated’ when B cells were loosely distributed within the synovium.

Rao D.A., Gurish M.F., Marshall J.L., Slowikowski K., Fonseka C.Y., Liu Y., Donlin L.T., Henderson L.A., Wei K., Mizoguchi F., Teslovich N.C., Weinblatt M.E., Massarotti E.M., Coblyn J.S., Helfgott S.M., Lee Y.C., Todd D.J., Bykerk V.P., Goodman S.M., Pernis A.B., Ivashkiv L.B., Karlson E.W., Nigrovic P.A., Filer A., Buckley C.D., Lederer J.A., Raychaudhuri S, & Brenner M.B. (2017). Pathologically expanded peripheral T helper cell subset drives B cells in rheumatoid arthritis. Nature, 542(7639), 110-114.

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