Sorting was performed on a FACS Aria II cell sorter (BD). Cells of interest were harvested from differentiation cultures, and resuspended in MACS buffer (5% FBS, Gibco; 0.5 mM EDTA in PBS). The following antibodies were used: CD41a-APC, CD235a-PE, CD43-PerCP, CD45-PerCP, CD34-FITC (BD Pharmingen), and CD56-APC (BioLegend). Dead cells were counterstained with Ghost Violet 540 cell viability dye (TONBO Biosciences). Sorting gates were set with appropriate Fluorescent Minus One (FMO) controls, and only live cells were collected.
Cd41a apc
Manufactured by BD
The CD41a-APC is a lab equipment product that detects the CD41a antigen using flow cytometry. It is a fluorescently labeled antibody that binds to the CD41a surface marker on cells. The CD41a-APC can be used to identify and analyze cell populations expressing the CD41a antigen.
Automatically generated - may contain errors
Lab products found in correlation
Cd235a pe, by BD (2 mentions)
Cd56 apc, by BioLegend (2 mentions)
Cd43 percp, by BD (2 mentions)
Cd34 fitc, by BD (2 mentions)
Cd45 percp, by BD (2 mentions)
Ghost violet 540 cell viability dye, by Cytek Biosciences (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Facsaria 2 cell sorter, by BD (1 mentions)
Cd45 pe, by BD (1 mentions)
Cd71 fitc, by BD (1 mentions)
Cd34 apc, by BD (1 mentions)
Cd117 apc, by Thermo Fisher Scientific (1 mentions)
Cd117 pe, by BD (1 mentions)
Tryple select, by Thermo Fisher Scientific (1 mentions)
Cd144 pe, by BD (1 mentions)
Kdr pe, by BD (1 mentions)
Streptavidin fitc, by BD (1 mentions)
Cd73 pe, by BD (1 mentions)
Facsaria 3 flow cytometer, by BD (1 mentions)
Clone hip8, by BD (1 mentions)
5 protocols using cd41a apc
1
Multiparameter Cell Sorting for Hematopoietic Lineages
2
Flow Cytometric Characterization of Adherent and Suspension Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Adherent cells were washed with Dulbecco's PBS (DPBS) (Thermo Fisher, Waltham, MA, #12563011) and treated with Tryp‐LE Select (Thermo Fisher, Waltham, MA, #12563011) at 37°C for 5 minutes, followed by gentle pipetting to complete cell dissociation. The dissociated cells were then washed with DPBS several times before antibody staining for flow cytometry. For suspension hematopoietic cells, gentle pipetting was used to dislodge any loosely adhered hematopoietic cells from the adherent stromal layer, and the cells were washed multiple times with PBS before staining. Cells were stained in ice‐cold fluorescence‐activated cell sorting buffer (PBS, 1% BSA). Antibodies used; KDR‐PE (BD Pharmingen, San Jose, CA, #560872), CD34‐APC (BD, San Jose, CA, #555824), CD144‐PE (BD, San Jose, CA, #561714), CD45‐PE (BD, San Jose, CA, #561866), CD117‐APC (Thermo Fisher, Waltham, MA, clone 104D2), CD117‐PE (BD, San Jose, CA, #555714), CD235a‐FITC/PE (BD, San Jose, CA, Clone GA‐R2 (HIR2)), CD41a‐APC (BD, San Jose, CA, #559777), CD71‐FITC (BD, San Jose, CA, #555536), CD73‐PE (BD, San Jose, CA, #561014), CXCR4‐Biotin (BD, San Jose, CA, #555973), and Streptavidin‐FITC (BD, San Jose, CA, #554060). Flow cytometry was conducted on a Stratadigm S1000EXI or Beckton Dickinson (BD) dual‐laser FacsCalibur flow cytometer. Analyses were conducted using FlowJo v8.7 (FlowJo, LLC) software.
3
Measuring Platelet Mitochondrial Potential
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mitochondrial membrane potential in isolated human platelets (200 × 106 ml−1) was measured using a flow cytometer FACSAria III (BD, Franklin Lakes, USA) with Diva version 7.0 acquisition and analysis software, using the probe TMRM (Life Techologies, Ref: T668), in non-quench mode (30 nM)19 (link) excited by 561 nM 40 mW laser and collected on 582/15 band pass filter. CD41a-APC (BD Pharmingen, Clone HIP8, Ref: 559777) at 18 times dilution was used to assess platelet activation. Samples were incubated with the probes in MiR05 for 30 min at room temperature. CI was inhibited using 2 μM rotenone. NV189 (250 μM) or DMSO control was added to the samples, followed by oligomycin (1 μg ml−1), FCCP (20 μM) and antimycin A (1 μg ml−1), the two latter additions as internal controls. Data software used was FlowJo 10 (Tree Star, Ashland, USA). Statistical analyses were performed, and all figures were generated using Prism 6 (GraphPad Software).
4
Platelet activation by leptospiral infection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After blood collection and PRP separation, samples were immediately processed for flow cytometry (FC). PRP (20 μL of 1 × 108 cells/mL solution) was mixed with the antibodies cocktail (20 μL in PBS, phosphate-buffered saline) and the test stimulus (40 μL in PBS). As stimuli, we used buffer only (negative control), 10 μM ADP (positive control for platelet activation) or L. interrogans Copenhageni (2 × 106 cells/mL). The reactions were incubated for 5, 10 or 30 min at RT, protected from light, then stopped and fixed by the addition of 500 μL 0.2% formaldehyde. The following conjugated antibodies dilutions were used: 5 μL CD41a-APC, 3 μL CD62P-BV421, 5 μL PAC-1-FITC, 5 μL CD63-FITC – all from BD Bioscience. Flow cytometric acquisition was performed with a FACS Canto II after optimization of settings using the cytometer setup and tracking beads. Instrument and reagents were from BD Bioscience. Data were analyzed using FlowJo version 7.6.5. The experiments were performed with four different blood donors, at least in duplicates. Data from 20,000 events in the CD41a-positive gate were collected for each sample.
5
Multi-Phenotypic Flow Cytometry Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Flow Cytometry was conducted using a MACSQuant® Analyzer 10 (Miltenyi Biotec), and the following antibodies: CD43-FITC(1G10), CD43-PerCP(1G10), CD235a-PE(HIR2), CD45-BV421(H130), CD45-PerCP(Hl30), CD94-Fitc(HP-3D9), CD38-FITC (HIT2), CD16APC(3G8), CD7-PE(MT701), CD16-FITC (3GB), CD42b-PE(HIP1), CD107a-FITC (H4A3) perforin-FITC (δG9), CD34 FITC (581), CD66BV421 (G10F5), CD16 PE (3G8), and CD14 PerCP (M5E2) from BD Pharmingen; CD41a-APC(REA386), CD45-APC(REA747), CD11b APC(REA713), CD201-PE (REA337), and anti-Ki67 (REA183) from Miltenyi Biotech, and CD56-PerCP(5.1H11), CD56-APC(5.1.H11), and CD45RA-APC (H1100) from BioLegend. All expansions were performed using ultra-low attachment surface, tissue culture plates (CORNING).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!