Platelets were isolated from mice 24 hours after injection with anti–CD42c-DyLight-x649 and were subsequently incubated with Fluo-4 AM (5 μM; Thermo Fisher Scientific) for 30 minutes at 37°C, followed by counterstaining with anti–CD41-BV421 (1:100; BD Biosciences) for 15 minutes and supplementation with calcium chloride (2 mM; Sigma). Baseline Fluo-4 fluorescence was recorded for 30 seconds, followed by challenge with PAR4-amide (25 μM), a collagen-related peptide (CRP-XL, 1 μg/mL; CAMBCOL), the stable thromboxane A2 mimetic U46619 (10 μM; Enzo), or ionomycin (10 μM; Invitrogen) and subsequent recording for 2 minutes. Samples were acquired on a BD LSRII (BD Bioscience) using FACSDiva acquisition software and analyzed using FlowJo software, version 10.
Anti cd41 bv421
Manufactured by BD
Anti-CD41-BV421 is a fluorescent-conjugated monoclonal antibody that binds to the CD41 antigen. CD41 is a glycoprotein found on the surface of platelets and megakaryocytes. The BV421 fluorophore is used for the detection and analysis of CD41-positive cells.
Automatically generated - may contain errors
Lab products found in correlation
Fluo 4 am, by Thermo Fisher Scientific (2 mentions)
Ionomycin, by Thermo Fisher Scientific (1 mentions)
U46619, by Enzo Life Sciences (1 mentions)
Calcium chloride, by Merck Group (1 mentions)
Lsr 2, by BD (1 mentions)
Fluoreporter mini biotin xx protein labeling kit, by Thermo Fisher Scientific (1 mentions)
Facsaria 2 sorter, by BD (1 mentions)
Fortessa, by BD (1 mentions)
Streptavidin apc, by BD (1 mentions)
2 protocols using anti cd41 bv421
1
Calcium Signaling in Isolated Murine Platelets
2
Isolation and Characterization of Pre-HSCs
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Cell suspensions were stained with following antibodies: Ter119-v500 clone (Clone TER-119); anti-CD41-BV421 (brilliant violet 421) or Alexa Fluor 488 (clone MW30reg); anti-CD45 FITC (clone 30-F11); biotinylated anti-VE-cadherin (clone 11.D4.1) followed by incubation with streptavidin-APC (all purchased from BD Pharmingen or BioLegend). Anti-mouse VE-cadherin (VC) antibody was biotinylated in-house using the FluoReporter Mini-Biotin-XX Protein Labeling Kit (Invitrogen). Cell populations were sorted using a FACSAria II sorter (BD) followed by purity checks. Type I pre-HSCs (Ter119−VC+CD41+CD45-) and type II pre-HSCs (Ter119−VC+CD41+CD45+) were sorted from E11 AGMs as described previously (Rybtsov et al., 2011 (link)). Dead cells were excluded using 7AAD staining and fluorescence minus one control staining was used to define gating strategies. Data acquisition and analyses were performed using Fortessa (BD) and FlowJo software.
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