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Streptavidin fitc conjugate

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Streptavidin-FITC conjugate is a protein complex consisting of the protein streptavidin covalently linked to the fluorescent dye FITC (fluorescein isothiocyanate). It is designed to bind to biotin-labeled biomolecules, allowing for their detection and visualization in various biological applications.

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Lab products found in correlation

Fish skin gelatin, by Merck Group (2 mentions) Biotinyl yvad cmk, by Enzo Life Sciences (2 mentions) Rabbit anti asc antibodies, by Adipogen (2 mentions) Alexa fluor 568 anti rabbit, by Thermo Fisher Scientific (2 mentions) Fixable viability dye, by Thermo Fisher Scientific (1 mentions) α cd45, by BioLegend (1 mentions) α cd4, by BioLegend (1 mentions) αcd56, by BioLegend (1 mentions) αcd19, by BioLegend (1 mentions) Facslyric, by BD (1 mentions) Facsverse, by BD (1 mentions)

3 protocols using streptavidin fitc conjugate

1

Siglec-7 Ligand Detection in AML Cells

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Detection of Siglec-7 ligands by flow cytometry was performed as previously described (6 (link)). AML-derived samples were analyzed as previously described (17 (link)). In brief, non-specific antibody binding by Fc receptors was blocked using 100 μL of Fc Receptor Blocker (Innovex Biosciences, Richmond, CA, USA) and dead cells were excluded from analysis by staining with a Fixable Viability Dye (ThermoFisher). For the Siglec ligands staining, recombinant human Siglec-7-hFc (R&D Systems, Minneapolis, MN, USA) was pre-incubated with PE-conjugated goat anti-human Ig (Jackson ImmunoResearch Laboratories, West Grove, USA) for 1 h at 4°C and then applied to the samples for 1 h at RT together with lineage-associated antibodies. Lineage-positive cells were stained with biotinylated αCD2, αCD14, αCD16, αCD19, αCD56, and αCD235, together with fluorophore-conjugated αCD8, αCD4, αCD45, αCD34, αCD38 (all from BioLegend), followed by a second step with streptavidin-FITC conjugate (BD Biosciences). Cells were washed and analyzed on a BD FACSVerse or BD FACSlyric (BD Biosciences). Data were analyzed using the FlowJo 10.0.6 software (Tree Star Inc.).
Haas Q., Markov N., Muerner L., Rubino V., Benjak A., Haubitz M., Baerlocher G.M., Ng C.K., Münz C., Riether C., Ochsenbein A.F., Simon H.U, & von Gunten S. (2022). Siglec-7 represents a glyco-immune checkpoint for non-exhausted effector memory CD8+ T cells with high functional and metabolic capacities. Frontiers in Immunology, 13, 996746.
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2

Quantitative ASC Aggregation Assay

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Primary monocyte-derived macrophages were differentiated on glass coverslips as described. Adherent macrophages were washed twice in PBS, primed with 10ng/mL LPS for 6 hours, followed by stimulation with 5µg/mL IC-FLA as noted. Cells were incubated with 2 µM biotinyl-YVAD-CMK (Enzo) during the last 4 hours of stimulation. Cells were then rinsed with PBS, fixed and permeabilized with ice cold methanol for 7 minutes at −20°C, and blocked in PBS containing 1% fish skin gelatin (Sigma), 2% Bovine Serum Albumin, 5% Normal Goat Serum for 1 h at room temperature (RT). ASC was detected with rabbit anti-ASC antibodies (Adipogen) in blocking buffer (one hour at RT). A streptavidin-FITC conjugate (BD biosciences) and anti-rabbit AlexaFluor 568 (Life Technologies) were used for secondary detection. Nuclei were counterstained with Hoechst 33342 and coverslips were mounted in Vectashield. Macrophages were analyzed on a Zeiss 780 confocal microscope using a 63× or 40× oil immersion objective. Quantitation of ASC aggregates was performed visually at 63× in at least 100 cells adjacently positioned along an arbitrary line.
Canna S.W., de Jesus A.A., Gouni S., Brooks S.R., Marrero B., Liu Y., DiMattia M.A., Zaal K.J., Montealegre Sanchez G.A., Kim H., Chapelle D., Plass N., Huang Y., Villarino A.V., Biancotto A., Fleisher T.A., Duncan J.A., O’Shea J.J., Benseler S., Grom A., Deng Z., Laxer R.M, & Goldbach-Mansky R. (2014). An activating NLRC4 inflammasome mutation causes autoinflammation with recurrent macrophage activation syndrome. Nature genetics, 46(10), 1140-1146.
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3

Quantitative ASC Aggregation Assay

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Primary monocyte-derived macrophages were differentiated on glass coverslips as described. Adherent macrophages were washed twice in PBS, primed with 10ng/mL LPS for 6 hours, followed by stimulation with 5µg/mL IC-FLA as noted. Cells were incubated with 2 µM biotinyl-YVAD-CMK (Enzo) during the last 4 hours of stimulation. Cells were then rinsed with PBS, fixed and permeabilized with ice cold methanol for 7 minutes at −20°C, and blocked in PBS containing 1% fish skin gelatin (Sigma), 2% Bovine Serum Albumin, 5% Normal Goat Serum for 1 h at room temperature (RT). ASC was detected with rabbit anti-ASC antibodies (Adipogen) in blocking buffer (one hour at RT). A streptavidin-FITC conjugate (BD biosciences) and anti-rabbit AlexaFluor 568 (Life Technologies) were used for secondary detection. Nuclei were counterstained with Hoechst 33342 and coverslips were mounted in Vectashield. Macrophages were analyzed on a Zeiss 780 confocal microscope using a 63× or 40× oil immersion objective. Quantitation of ASC aggregates was performed visually at 63× in at least 100 cells adjacently positioned along an arbitrary line.
Canna S.W., de Jesus A.A., Gouni S., Brooks S.R., Marrero B., Liu Y., DiMattia M.A., Zaal K.J., Montealegre Sanchez G.A., Kim H., Chapelle D., Plass N., Huang Y., Villarino A.V., Biancotto A., Fleisher T.A., Duncan J.A., O’Shea J.J., Benseler S., Grom A., Deng Z., Laxer R.M, & Goldbach-Mansky R. (2014). An activating NLRC4 inflammasome mutation causes autoinflammation with recurrent macrophage activation syndrome. Nature genetics, 46(10), 1140-1146.
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