Rea1047

Manufactured by Miltenyi Biotec

The REA1047 is a lab equipment product from Miltenyi Biotec. It is a device used for cell separation and purification. The core function of the REA1047 is to isolate specific cell types from complex samples, such as blood or tissue, through magnetic separation techniques.

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Lab products found in correlation

Fib27, by BioLegend (1 mentions) Facsaria 2 sorp, by BD (1 mentions) Cd45ra, by BioLegend (1 mentions) Viogreen, by Miltenyi Biotec (1 mentions) Hit3a, by BioLegend (1 mentions) Apc vio770, by Miltenyi Biotec (1 mentions) Efluor 780, by Thermo Fisher Scientific (1 mentions) Cxcr5 brilliant violet 421, by BioLegend (1 mentions) Facsaria fusion, by BD (1 mentions) Facscanto 2, by BD (1 mentions) Fitc annexin 5 apoptosis detection kit 1, by BD (1 mentions)

2 protocols using rea1047

Isolation and Characterization of Memory T Cells

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FACS was performed to isolate α4⁺ memory T cells for subsequent scRNASeq, with α4⁺ memory T cells being defined as CD3⁺CD45RA⁻α4⁺ viable single lymphocytes. To that end, the following fluorescently-labeled antibodies were used in combination with fixable viability dye eFluor780 (eBioscience (1:1000)): CD3 (APC, HIT3a, Biolegend (1:100)), α4 (PE/Cy7, 9F10, Biolegend (1:100)), CD45RA (VioGreen, REA1047, Miltenyi Biotec (1:100)).
To isolate β7^+/− CD4 (CD3⁺TCRVα7.2⁻CD4⁺CD8a⁻CD45RA⁻ β7^+/− single lymphocytes) and CD8 (CD3⁺TCRVα7.2⁻CD4⁻CD8a⁺CD45RA⁻ β7^+/− single lymphocytes) memory T cells for functional investigations and CD103-inducibility analysis, we used fluorescently-labeled antibodies against the following epitopes: CD3 (PE/Cy7, SK7, Biolegend (1:1000)), TCRVα7.2 (BV421, OF5A12, BD Biosciences (1:500)), CD4 (APC-Vio770, VIT4, Miltenyi Biotec (1:2000)), CD8a (FITC, RPA-T8, Biolegend (1:2000)), CD45RA (VioGreen, REA1047, Miltenyi Biotec (1:1000)), β7 (PE, FIB27, Biolegend (1:1000)).
FACS Aria II SORP (BD Biosciences) instruments were used for FACS.
MACS enrichment for CD4 and CD8 T cells from PBMCs was performed according to manufacturer’s instructions using the CD4 T Cell Isolation Kit, human and CD8 T Cell Isolation Kit, human, respectively (both Miltenyi Biotec).

Wiendl M., Dedden M., Liu L.J., Schweda A., Paap E.M., Ullrich K.A., Hartmann L., Wieser L., Vitali F., Atreya I., Müller T.M., Günther C., Atreya R., Neurath M.F, & Zundler S. (2024). Etrolizumab-s fails to control E-Cadherin-dependent co-stimulation of highly activated cytotoxic T cells. Nature Communications, 15, 1043.

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CVID Patient PBMC Apoptosis Assay

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PBMC isolated from CVID patients and HC were stained with surface antibodies against CD45RA‐FITC (REA1047, Miltenyi Biotec), CD3‐PE (SK7, BD bioscience), CD4‐PO (VIT4, Miltenyi Biotec), CXCR5‐Brilliant Violet 421 (J252D4, BioLegend), CD25‐APC (BC96, BioLegend) (Supporting Information Table S4, staining panel D) and FACS‐sorted into cTfh (CD4⁺CD45RA⁻CXCR5⁻), T naïve (CD4⁺CD45RA⁺), and Tconv (CD4⁺CD45RA⁻CXCR5⁻) (FACS Aria Fusion [BD]). cTfh, T naïve, and Tconv, once isolated, were seeded in a 96‐wells plate, activated with anti‐CD3/CD28 beads (1:3 ratio, cells:beads) in complete medium (RPMI 10%FBS, PS/G) and incubated for 4 days at 37°C. At the end of incubation, cells were stained with Annexin V/propidium iodine (PI) following the FITC Annexin‐V Apoptosis Detection Kit I BD Pharmingen (Cat. 556547) protocol (Supporting Information Table S4, staining panel E). Cells were acquired on FACS CantoII (BD) and analyzed with FlowJo (Tree Star) software.

Milardi G., Di Lorenzo B., Gerosa J., Barzaghi F., Di Matteo G., Omrani M., Jofra T., Merelli I., Barcella M., Filippini M., Conti A., Ferrua F., Pozzo Giuffrida F., Dionisio F., Rovere‐Querini P., Marktel S., Assanelli A., Piemontese S., Brigida I., Zoccolillo M., Cirillo E., Giardino G., Danieli M.G., Specchia F., Pacillo L., Di Cesare S., Giancotta C., Romano F., Matarese A., Chetta A.A., Trimarchi M., Laurenzi A., De Pellegrin M., Darin S., Montin D., Marinoni M., Dellepiane R.M., Sordi V., Lougaris V., Vacca A., Melzi R., Nano R., Azzari C., Bongiovanni L., Pignata C., Cancrini C., Plebani A., Piemonti L., Petrovas C., Di Micco R., Ponzoni M., Aiuti A., Cicalese M.P, & Fousteri G. (2022). Follicular helper T cell signature of replicative exhaustion, apoptosis, and senescence in common variable immunodeficiency. European Journal of Immunology, 52(7), 1171-1189.

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