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Cd41a

Manufactured by BD
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Sourced in United States, Denmark

The CD41a is a laboratory equipment product manufactured by BD. It is designed to measure and analyze certain types of cells within a biological sample. The core function of the CD41a is to detect and quantify the presence of specific cell surface markers, providing information about the cellular composition of the sample. The detailed specifications and intended applications of this product are not available for this response.

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Lab products found in correlation

Cd41a, by Novus Biologicals (2 mentions) One micrometer beads, by Thermo Fisher Scientific (2 mentions) Anti e selectin, by Novus Biologicals (2 mentions) Annexin 5, by BD (1 mentions) Cellgro media, by CellGenix (1 mentions) Pe positive selection kit, by STEMCELL (1 mentions) Il 1b, by R&D Systems (1 mentions) Facsclibur, by BD (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Iscove s modified dulbecco s medium, by Thermo Fisher Scientific (1 mentions) B27 without vitamin a, by Thermo Fisher Scientific (1 mentions) Ham s f12, by Corning (1 mentions) β tubulin, by Abcam (1 mentions) Lsm 880 confocal microscope, by Zeiss (1 mentions) Accucount ultra rainbow fluorescent particles, by Spherotech (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Beckman Coulter (1 mentions) 7 aad, by Beckman Coulter (1 mentions)

Spelling variants of this product

CD41a-APCH7 (5 citations) CD41a-PE (4 citations) Anti-CD41a-FITC (3 citations) FITC-conjugated anti-CD41a antibody (3 citations) Anti-human CD41a PE (2 citations) Anti-human CD41a antibody (2 citations) CD41a- PE-Cy7 (clone HIP8, (2 citations) CD41a (HIP8) (2 citations) PE-conjugated anti-CD41a (2 citations) APC-conjugated CD41a (1 citations)

7 protocols using cd41a

1

Characterization of Blood Microparticles

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Microparticles were analyzed in the blood. Plasma (250 μlL aliquots) was centrifuged at 20,000 x g for 2.5 hours. The pellet was re-suspended in Hank’s buffered saline solution containing 20 mM HEPES and 5 mM glucose. It was re-suspended by vortexing the pellet for 1.5 minute in buffer at a volume of 40% of the initial plasma volume. Microparticles may be distinguished based upon protein, lipid and cholesterol composition. Multi-color fluorescence-activated cell sorting was performed to characterize the cellular source and activation state of the microparticles. An aliquot of microparticles was stained with antibodies to CD41a (Novus Biologicals) and CD31 (BD Biosciences) to identify whether they originated from platelets (CD41a+, CD31+) or endothelial cells (CD41a−CD31+). One micrometer beads (Molecular Probes) were used to gate the particles based on size. The number of microparticles from each cell type (platelet or endothelial) was measured in a 250 μl plasma sample. We measured CD62E levels (anti-E-selectin, Novus Biologicals) on the surface of the endothelial cell microparticles as an indicator of whether the cells that released the particles were activated.
Kataria A., Levine D., Wertenteil S., Vento S., Xue J., Rajendiran K., Kannan K., Thurman J.M., Morrison D., Brody R., Urbina E., Attina T., Trasande L, & Trachtman H. (2017). Exposure to Bisphenols and Phthalates and Association with Oxidant Stress, InsulinN Resistance, and Endothelial Dysfunction in Children. Pediatric research, 81(6), 857-864.
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2

Characterization of Blood Microparticles

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Microparticles were analyzed in the blood. Plasma (250 μlL aliquots) was centrifuged at 20,000 x g for 2.5 hours. The pellet was re-suspended in Hank’s buffered saline solution containing 20 mM HEPES and 5 mM glucose. It was re-suspended by vortexing the pellet for 1.5 minute in buffer at a volume of 40% of the initial plasma volume. Microparticles may be distinguished based upon protein, lipid and cholesterol composition. Multi-color fluorescence-activated cell sorting was performed to characterize the cellular source and activation state of the microparticles. An aliquot of microparticles was stained with antibodies to CD41a (Novus Biologicals) and CD31 (BD Biosciences) to identify whether they originated from platelets (CD41a+, CD31+) or endothelial cells (CD41a−CD31+). One micrometer beads (Molecular Probes) were used to gate the particles based on size. The number of microparticles from each cell type (platelet or endothelial) was measured in a 250 μl plasma sample. We measured CD62E levels (anti-E-selectin, Novus Biologicals) on the surface of the endothelial cell microparticles as an indicator of whether the cells that released the particles were activated.
Kataria A., Levine D., Wertenteil S., Vento S., Xue J., Rajendiran K., Kannan K., Thurman J.M., Morrison D., Brody R., Urbina E., Attina T., Trasande L, & Trachtman H. (2017). Exposure to Bisphenols and Phthalates and Association with Oxidant Stress, InsulinN Resistance, and Endothelial Dysfunction in Children. Pediatric research, 81(6), 857-864.
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3

Isolation and Characterization of Plasma Microparticles

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Plasma MPs were isolated from platelet-poor plasma by sequential centrifugations, as described previously.12 (link) MP-free plasma was collected and pelleted MPs were washed and re-suspended in RPMI 1640 (Hyclone). MP surface markers were analysed by flow cytometry, using antibodies against annexin V (BD Biosciences, 556547, for total MPs), CD31 (BD Biosciences, 555448, for endothelial MPs), CD41a (BD Biosciences, 555467, for platelet MPs), CD235a (BD Biosciences, 340947, for red blood cell MPs) and CD45 (BD Biosciences, 347464, for leukocyte MPs), as described previously.12 (link)
Hu Y., Li H., Yan R., Wang C., Wang Y., Zhang C., Liu M., Zhou T., Zhu W., Zhang H., Dong N, & Wu Q. (2018). Increased Neutrophil Activation and Plasma DNA Levels in Patients with Pre-Eclampsia. Thrombosis and haemostasis, 118(12), 2064-2073.
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4

Isolation and Enrichment of Megakaryocytes

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CD34+ progenitor cells were isolated and enriched from cord blood as reported previously.(51) MK cultures of 105 cells/ml were supplemented with 100 ng/ml thrombopoietin (rhTPO CellGenix, Freiburg, Germany) and 10 ng/ml IL1b (R&D, Minneapolis, MN, USA) in CellGro media (CellGenix, Freiburg, Germany). Media and cytokines were rejuvenated at days 3 and 6. At day 10, mature MKs were immunopurified using an anti-CD42b PE conjugated antibody (Pab5, NHS Blood and Transplant, IBGRL, Bristol, England) and a PE positive selection kit (STEMCELL Technologies, Vancouver, Canada). MK purity was verified by FACS and shown to be >95% for CD41a (BD, San Jose, CA, USA) and CD42b (IBGRL, Bristol, England).
Zou S., Teixeira A.M., Kostadima M., Astle W.J., Radhakrishnan A., Simon L.M., Truman L., Fang J.S., Hwa J., Zhang P.X., van der Harst P., Bray P.F., Ouwehand W.H., Frontini M, & Krause D.S. (2017). SNP in human ARHGEF3 promoter is associated with DNase hypersensitivity, transcript level and platelet function, and Arhgef3 KO mice have increased mean platelet volume. PLoS ONE, 12(5), e0178095.
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5

Flow Cytometric Analysis of Platelet Activation

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After inhibition of in vitro platelet activation with prostagladin E1, 5 μL of the samples were stained with 1 µl of a fluorescein isothiocyanate (FITC) conjugated monoclonal antibody against GPIIbIIIa (CD41a, BD Biosciences CA), 1 µl of a phycoerythrin (PE)-conjugated monoclonal antibody against P-selectin (CD62P, BD Biosciences, CA) and 1 µl of a allophycocyanin (APC)-conjugated monoclonal antibody against glycophorin A (GPA, Dakopatts Glostrup, Denmark). The mixtures were incubated for 15 min at room temperature. 150 µL of 1xPBS, pH 7.4 was added to the stained platelets which were immediately analyzed by flow cytometry (FACsClibur, BD Biosciences, San Jose, CA) as described elsewhere42 (link).
Chanpeng P., Svasti S., Paiboonsukwong K., Smith D.R, & Leecharoenkiat K. (2019). Platelet proteome reveals specific proteins associated with platelet activation and the hypercoagulable state in β-thalassmia/HbE patients. Scientific Reports, 9, 6059.
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6

Megakaryocyte Differentiation from iPSC-derived HPCs

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Human iPSCs were differentiated into HPCs using the previously described protocol [19 (link)]. Cells were plated on Matrigel for differentiation and the medium and cytokine changes were followed as described. Cells were cultured at 37°C with 4% O2/5% CO2 for 9 days, and HPCs were collected. Cells were analyzed by flow cytometry to confirm the surface expression of CD41a and CD235a (BD Bioscience). The HPCs were further differentiated to MKs in serum-free differentiation medium, Iscove’s Modified Dulbecco’s Medium (Thermo Fisher Scientific) containing 25% Ham’s F-12 (Corning), 0.5% N2 (Thermo Fisher Scientific), 1% B27 without vitamin A (Thermo Fisher Scientific), 0.05% bovine serum albumin (BSA; Sigma), 2 mM l-glutamine, and penicillin /streptomycin supplemented with 50 ng/mL SCF and 50 ng/mL TPO at 37°C, 5% CO2 for 6–7 days. Tamoxifen was added at day 0 of MK differentiation for 24 h to disrupt Lyn unless stated otherwise. HPC proliferation was analyzed by counting cell number from day 0–6 of MK differentiation.
Moroi A.J, & Newman P.J. (2021). Conditional CRISPR-mediated deletion of Lyn kinase enhances differentiation and function of iPSC-derived megakaryocytes. Journal of thrombosis and haemostasis : JTH, 20(1), 182-195.
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7

Comprehensive Platelet Analysis by Flow Cytometry

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Collected particles, whole blood, and washed platelets were analyzed using flow cytometry. Half of each sample was activated with 3U/mL thrombin for 5 minutes. All samples were fixed in 3.7% formaldehyde, washed in FACS buffer (2% fetal bovine serum in PBS), and stained for CD41a, IgG (BD Biosciences), DAPI, and 7AAD (Beckman Coulter) for 30 minutes at RT. The cells were washed and analyzed on FACS CANTO2. The number of particles collected from each vessel was calculated using AccuCount Ultra Rainbow Fluorescent Particles (Spherotech). Analysis was performed on FLOWJO. Fixed quiescent and activated particles were permeabilized with Triton X-100, stained with β-tubulin (1:100, Abcam) overnight, washed, and incubated with Alexa Fluor 488 for 1 hour. Particles were washed, resuspended into a 1% agarose solution, mounted on coverslips, and imaged with a Zeiss LSM 880 confocal microscope.
Kotha S., Sun S., Adams A., Hayes B., Phong K.T., Nagao R., Reems J.A., Gao D., Torok-Storb B., López J.A, & Zheng Y. (2018). Microvasculature-directed thrombopoiesis in a 3D in vitro marrow microenvironment. PLoS ONE, 13(4), e0195082.
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