PE-conjugated rat anti-human CD184/CXCR4 (clone 1D9, isotype control rat IgG2a,κ), FITC-conjugated mouse anti-Rac1 (clone 102/Rac1, isotype control mouse IgG2,b), APC-conjugated mouse anti-human CD34 (clone 581, isotype mouse IgG1,κ), PE-conjugated mouse anti-human CD38 (clone HIT2, isotype control mouse IgG1,κ) and APC-conjugated mouse anti-human CD45 (clone Hl30, isotype mouse IgG1,κ) were purchased from BD Biosciences (San Diego, CA, USA). Blocking reagents human gamma globulin and mouse gamma globulin were purchased from Jackson ImmunoResearch Laboratories Incorporated (West Grove, PA, USA). BD Cytofix™ fixation buffer was purchased from BD Biosciences. Recombinant human SDF-1α was purchased from R&D Systems. FITC-conjugated Cholera toxin B subunit (CTxB) and methyl-β-cyclodextrin (MβCD) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Rac1 inhibitor NSC23766 was purchased from BioVision (Milpitas, CA, USA).
Fitc conjugated cholera toxin b subunit ctxb
Manufactured by Merck Group
Sourced in United States
FITC-conjugated Cholera toxin B subunit (CTxB) is a fluorescently labeled protein used as a cell membrane marker. It binds to the GM1 ganglioside, a glycosphingolipid found on the surface of many cell types. The FITC (Fluorescein isothiocyanate) label allows visualization of the bound CTxB under fluorescent microscopy.
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2 protocols using fitc conjugated cholera toxin b subunit ctxb
1
Immunophenotyping of Hematopoietic Stem Cells
2
Membrane Fusion Assay for S. pneumoniae EVs
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To assess membrane fusion using S. pneumoniae EVs and J774A.1 macrophage cells, we essentially followed procedures described earlier [19 (link)]. Briefly, EVs were labeled with 1 mg/mL rhodamine isothiocyanate B-R18 (Molecular Probes) for 1 h at room temperature. An unlabeled probe was removed by centrifugation at 100,000× g (60 min, 4 °C). After washing with PBS, labeled EVs were resuspended in 1 mL PBS. Subsequently, the host cell plasma membrane was labeled for 1 h prior to the incubation with EVs with 8 mg/mL FITC-conjugated cholera toxin B subunit (CtxB) (Sigma-Aldrich). Then, labeled EVs were added in a 1:4 dilution in the wells and incubated for 30 min at 37 °C. When applicable, 10 mg/mL Filipin III was added 30 min prior to the addition of EVs. After incubation with EVs, cell samples were analyzed by confocal microscopy as described above.
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