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Cd90 apc clone 5e10

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CD90-APC clone 5E10 is a fluorochrome-conjugated monoclonal antibody that specifically binds to the CD90 (Thy-1) surface antigen. CD90 is a glycosylphosphatidylinositol (GPI)-anchored glycoprotein expressed on various cell types, including thymocytes, neurons, and mesenchymal stem cells.

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Lab products found in correlation

Cd9 apc clone hi9a, by BioLegend (3 mentions) Cd63 apc clone h5c6, by BioLegend (3 mentions) Cd81 apc clone 5a6, by BioLegend (3 mentions) Cd44 apc clone bj18, by BioLegend (3 mentions) Cd73 apc clone ad2, by BioLegend (3 mentions) Cfda se, by Merck Group (2 mentions) Cfda se, by BioLegend (2 mentions) Fitc fluorescent nanobeads, by Biocytex (2 mentions) Cd34 bv785 clone 561, by BioLegend (1 mentions) Y 27632, by AbMole (1 mentions) Moflow astrios, by Beckman Coulter (1 mentions) Facsaria fusion sorter, by BD (1 mentions) Tryple express, by Thermo Fisher Scientific (1 mentions) E cadherin clone eccd 2, by Thermo Fisher Scientific (1 mentions) Lsm 880 microscope, by Zeiss (1 mentions)

Close spelling variants of this product

APC/Cy7-CD90 (clone 5E10, APC anti-human CD90 (clone 5E10, APC-CD90 (5E10, CD90-APC

6 protocols using cd90 apc clone 5e10

1

Cynomolgus Macaque Hematopoietic Stem Cell Quantification

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Cynomolgus macaques37 (link) were treated with intravenous injection of CD117-ADC (0.1 or 0.3 mg/kg × 1 day) or busulfan (6 mg/kg × 4 days), and bone marrow CD34 + CD90 + CD45RA- cells were quantified by using antibodies against CD34 BV785 (clone 561, BioLegend, 1:300), CD90 APC (clone 5E10, BioLegend, 1:300), and CD45RA VioBlue (clone T6D11, Miltenyi Biotec, 1:50) in flow cytometry 7 days after drug administration (n = 3 per group) and compared to historical phosphate-buffered saline (PBS)-treated naïve animals. Bone marrow cellularity was assessed by complete blood count analysis to determine white blood cells per ml of marrow. Flow cytometry was used to determine the frequency of CD34 + CD90 + CD45RA- cells of total CD45+ cells. The absolute number of CD34 + CD90 + CD45RA- was determined by multiplying the frequency of CD34 + CD90 + CD45RA- cells (out of total CD45 cells) with the bone marrow cellularity in cells per ml.
Uchida N., Stasula U., Demirci S., Germino-Watnick P., Hinds M., Le A., Chu R., Berg A., Liu X., Su L., Wu X., Krouse A.E., Linde N.S., Bonifacino A., Hong S.G., Dunbar C.E., Lanieri L., Bhat A., Palchaudhuri R., Bennet B., Hoban M., Bertelsen K., Olson L.M., Donahue R.E, & Tisdale J.F. (2023). Fertility-preserving myeloablative conditioning using single-dose CD117 antibody-drug conjugate in a rhesus gene therapy model. Nature Communications, 14, 6291.
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2

Single-Cell Sorting of Organoid Populations

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Organoids were dissociated into single-cell suspensions using TrypLE Express (ThermoFisher) supplemented with Rho-kinase inhibitor Y-27632 (10 µM, Abmole). Single-cell suspensions were stained using mouse Alexa-fluor 488 anti-human CD326 EPCAM clone 9C4 (BioLegend, 324210, 1:20), CD90-APC clone 5E10 (BioLegend, 328113, 1:50) as described71 (link). Populations were sorted using BD FACSAria—Fusion sorter (BD Biosciences) or MoFlow® Astrios (Beckman Coulter) and used for their respective applications (Supplementray Fig. 13). Data were analysed with software Kaluza analysis v2.1.
Calandrini C., Schutgens F., Oka R., Margaritis T., Candelli T., Mathijsen L., Ammerlaan C., van Ineveld R.L., Derakhshan S., de Haan S., Dolman E., Lijnzaad P., Custers L., Begthel H., Kerstens H.H., Visser L.L., Rookmaaker M., Verhaar M., Tytgat G.A., Kemmeren P., de Krijger R.R., Al-Saadi R., Pritchard-Jones K., Kool M., Rios A.C., van den Heuvel-Eibrink M.M., Molenaar J.J., van Boxtel R., Holstege F.C., Clevers H, & Drost J. (2020). An organoid biobank for childhood kidney cancers that captures disease and tissue heterogeneity. Nature Communications, 11, 1310.
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3

Characterization of Extracellular Vesicles

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Flow cytometry: Cleared secretome was 1:1 diluted with PBS and divided into 3 aliquots: (i) unstained, (ii) CFSE (1 µM final concentration) stained for 30 min at 37 °C and (iii) after CFSE supplementation, further staining for 30 min at 4 °C with one of the following antibodies: CD9-APC clone HI9A, CD63-APC clone H5C6, CD81-APC clone 5A6, CD44-APC clone BJ18, CD73-APC clone AD2 or CD90-APC clone 5E10) (Biolegend, San Die-go, CA, USA). After a further 1:3 dilution with PBS, samples were analyzed with a CytoFlex flow cytometer. FITC-fluorescent beads of 160, 200, 240 and 500 nm (Biocytex, Mar-seille, France) were used as an internal control. At least 30,000 events were collected.
Nanoparticle tracking analysis (NTA): Cleared secretome was 1:1 diluted in PBS and visualized by Nanosight LM10-HS system (NanoSight Ltd., Amesbury, UK). Each sample was run with 5 recordings of 60 s. NTA software provided both concentration measurements and high-resolution distribution profiles of particle size.
Ragni E., Perucca Orfei C, & de Girolamo L. (2022). Secreted Factors and Extracellular Vesicles Account for the Immunomodulatory and Tissue Regenerative Properties of Bone-Marrow-Derived Mesenchymal Stromal Cells for Osteoarthritis. Cells, 11(21), 3501.
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4

3D Imaging of Organoid Structures

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High-resolution 3D imaging on organoids was performed as described40 (link) using the following antibodies: SIX2 (Proteintech, 11562-1-AP, 1:200), E-cadherin clone ECCD-2 (ThermoFisher, 13-1900, 1:500), CD90-APC clone 5E10 (BioLegend, 328113, 1:200). Imaging was performed using Zeiss LSM880 microscope. Three-dimensional reconstruction was performed using the software Imaris v.9.2.1.
Calandrini C., Schutgens F., Oka R., Margaritis T., Candelli T., Mathijsen L., Ammerlaan C., van Ineveld R.L., Derakhshan S., de Haan S., Dolman E., Lijnzaad P., Custers L., Begthel H., Kerstens H.H., Visser L.L., Rookmaaker M., Verhaar M., Tytgat G.A., Kemmeren P., de Krijger R.R., Al-Saadi R., Pritchard-Jones K., Kool M., Rios A.C., van den Heuvel-Eibrink M.M., Molenaar J.J., van Boxtel R., Holstege F.C., Clevers H, & Drost J. (2020). An organoid biobank for childhood kidney cancers that captures disease and tissue heterogeneity. Nature Communications, 11, 1310.
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5

Comprehensive Characterization of Extracellular Vesicles

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All analyses were performed after 1:1 secretome dilution with PBS.
Nanoparticle tracking analysis (NTA): secretomes were run by Nanosight NS-300 system (NanoSight Ltd., Amesbury, UK) (5 recordings of 60 s) and EVs visualized with NTA software v3.4 providing both high-resolution particle size distribution profiles and concentration measurements.
Flow cytometry: 3 aliquots were analyzed. (i) Unstained, (ii) 5(6)-carboxyfluorescein diacetate succinimidyl ester (CFDA-SE, Sigma-Aldrich, St. Louis, MO, USA) stained (1 µM final concentration, 30 min at 37 °C) to visualize EVs after transformation into FITC-channel fluorescent carboxyfluorescein succinimidyl ester (CFSE), iii) after CFDA-SE supplementation, stained (30 min at 4 °C) with CD9-APC clone HI9A, CD63-APC clone H5C6, CD81-APC clone 5A6, CD44-APC clone BJ18, CD73-APC clone AD2, CD90-APC clone 5E10 (Biolegend, San Die-go, CA, USA). Samples were analyzed with a CytoFlex flow cytometer collecting at least 30,000 events. FITC-fluorescent nanobeads (160, 200, 240, and 500 nm, Biocytex, Marseille, France) were used as internal control for efficient detection in the nanometric range.
Ragni E., Perucca Orfei C., Valli F., Zagra L, & de Girolamo L. (2022). Molecular Characterization of Secreted Factors and Extracellular Vesicles-Embedded miRNAs from Bone Marrow-Derived Mesenchymal Stromal Cells in Presence of Synovial Fluid from Osteoarthritis Patients. Biology, 11(11), 1632.
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6

Characterizing Extracellular Vesicle Populations

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Flow cytometry: cleared secretomes were 1:1 diluted with PBS and divided into 3 aliquots: (i) unstained, (ii) 5(6)-carboxyfluorescein-diacetate-succinimidyl-ester (CFDA-SE, Sigma-Aldrich, St. Louis, MO, USA)-stained (1 µM final concentration, 30 min at 37 °C), (iii) after CFDA-SE supplementation and incorporation leading to FITC-fluorescent carboxyfluorescein succinimidyl ester (CFSE), CD9-APC clone HI9A, CD63-APC clone H5C6, CD81-APC clone 5A6, CD44-APC clone BJ18, CD73-APC clone AD2, CD90-APC clone 5E10 (Biolegend, San Diego, CA, USA) stained (30 min at 4 °C). After a further 1:3 dilution with PBS, samples were analyzed with a CytoFlex flow cytometer. At least 30,000 events were collected. FITC-fluorescent nanobeads of 160, 200, 240, and 500 nm (Biocytex, Marseille, France) were used as internal control.
Nanoparticle tracking analysis (NTA): cleared secretomes were 1:1 diluted in PBS and visualized by Nanosight NS-300 system (NanoSight Ltd., Amesbury, UK) (5 recordings of 60 s). NTA software v3.4 provided both concentration measurements and high-resolution particle size distribution profiles.
Ragni E., Perucca Orfei C., De Luca P., Libonati F, & de Girolamo L. (2022). Tissue-Protective and Anti-Inflammatory Landmark of PRP-Treated Mesenchymal Stromal Cells Secretome for Osteoarthritis. International Journal of Molecular Sciences, 23(24), 15908.
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