Rea293

Manufactured by Miltenyi Biotec

Sourced in Germany

The REA293 is a laboratory equipment product offered by Miltenyi Biotec. It serves as a core function to support research and analysis activities in relevant scientific fields.

Automatically generated - may contain errors

Lab products found in correlation

9 protocols using rea293

Cytotoxic NK Cell Receptor Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

For the analysis of activating receptor expression by cytotoxic NK cells (CD3-; CD56dim), 5 × 10⁶ human PBMCs were treated with the respective chemotherapeutic compound and incubated for 72 h (37 °C/5% CO₂). Then cells were harvested and 1 × 10⁶ live cells were washed with wash buffer, followed by incubation with 10 µL of Tandem Signal Enhancer (Miltenyi Biotec). Incubation with the following antibodies in a total volume of 100 µL was conducted for 20 min at RT: CD3-VioGreen (REA613, 1:200), CD56-APC-Vio770 (REA196, 1:200), CD226-VioBlue (REA1040, 1:50); CD335 (NKp46)-Vio Bright B515 (REA808, 1:50), CD337 (NKp30)-PE (REA823, 1:75), CD336 (NKp44)-APC (REA1163, 1:75), and CD314 (NKG2D)-PE-Vio 770 (REA1228, 1:75), all from Miltenyi Biotec. As isotype controls served respective REA controls (Miltenyi Biotec, REA293). To exclude dead cells from analysis, 0.25 µg propidium iodide solution was added prior to acquisition. At least 20,000 CD3−/CD56+ cells were analyzed for each sample.

Troschke-Meurer S., Zumpe M., Meißner L., Siebert N., Grabarczyk P., Forkel H., Maletzki C., Bekeschus S, & Lode H.N. (2023). Chemotherapeutics Used for High-Risk Neuroblastoma Therapy Improve the Efficacy of Anti-GD2 Antibody Dinutuximab Beta in Preclinical Spheroid Models. Cancers, 15(3), 904.

+ Open protocol

+ Expand

Evaluation of AKTpS473 in Diagnostic Blast Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Diagnostic blast cells were obtained from thawed cryopreserved BM samples after red blood cell lysis. Fixation, permeabilization and staining (with both intracellular and cell surface markers) were performed using the PerFix-no centrifuge assay kit (Beckman Coulter) according to the manufacturer’s instructions. The antibody panel contained: anti-AKTpS473-Vio515 (clone: REA359, Miltenyi Biotec), anti-CD33-PC5.5 (clone D3HL60.251, Beckman Coulter), anti-CD34-PC7 (clone 581, Beckman Coulter), anti-CD117/KIT-APC (clone 104D2D1, Beckman Coulter), anti-CD3-AA750 (clone UCHT1, Beckman Coulter), anti-CD4-PB (clone 13B8.2, Beckman Coulter) and anti-CD45-KO (clone J33, Beckman Coulter). Blast cells were gated as CD45^dim, SSC^low, CD33+, excluding lymphocytes (CD45^bright, SSC^low, CD33−), monocytes (CD45int/bright, SSC^int, CD33^bright) and mature myelomonocytic cells (CD45^int, SSC^high, CD33^dim/neg). Isotype control (clone REA293, Miltenyi Biotec) was used to better define the threshold of AKTpS473-positive cells. AKTpS473 expression levels were calculated as [mean fluorescent intensity (MFI) of blast cells/MFI of isotype IgG control]. Measurements were performed on a Navios flow cytometer and analyzed with Kaluza software (Beckman-Coulter).

Duployez N., Boudry-Labis E., Roumier C., Boissel N., Petit A., Geffroy S., Helevaut N., Celli-Lebras K., Terré C., Fenneteau O., Cuccuini W., Luquet I., Lapillonne H., Lacombe C., Cornillet P., Ifrah N., Dombret H., Leverger G., Jourdan E, & Preudhomme C. (2018). SNP-array lesions in core binding factor acute myeloid leukemia. Oncotarget, 9(5), 6478-6489.

+ Open protocol

+ Expand

Cardiomyocyte Differentiation and Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

On day 27 ± 2 of the differentiation the cells were detached and dissociated with Detachment Kit 2 (Promocell, Heidelberg, Germany) centrifuged 5 min at 200 g then fixed with paraformaldehyde (4%) for 10 min at RT (Sigma-Aldrich) and permeabilized with 90% cold methanol for 15 min at +4 °C . The cells were washed 3 times with PBS and then stained with Anti-Cardiac Troponin T-APC 1:100, recombinant human IgG1, clone REA400 (MiltenyiBiotec, Bergisch Gladbach, Germany) or CTL-I APC 1:100, Monoclonal REA Control (I) antibody human, clone REA293 (MiltenyiBiotec) diluted in PBS plus 0.1% Triton X-100 and 0.5% BSA for 45min at RT in the dark. The cells were washed and resuspended with PBS plus 0.5% BSA and collected on MACSQuant^® Analyzer 10 (MiltenyiBiotec) and analyzed using FlowJo.

Jeziorowska D., Fontaine V., Jouve C., Villard E., Dussaud S., Akbar D., Letang V., Cervello P., Itier J.M., Pruniaux M.P, & Hulot J.S. (2017). Differential Sarcomere and Electrophysiological Maturation of Human iPSC-Derived Cardiac Myocytes in Monolayer vs. Aggregation-Based Differentiation Protocols. International Journal of Molecular Sciences, 18(6), 1173.

+ Open protocol

+ Expand

Cytochrome-c Release Assay in Macrophages

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cytochrome‐c release was measured under relevant specified conditions (see figure legends for details), as performed previously (Waterhouse & Trapani, 2003 (link)). Briefly, either unstained BMMo or macrophages, or Ly6C⁺‐stained monocytes were washed 1× in PBS and resuspended in 50 μl MELB buffer (20 mM HEPES pH 7.5, 100 mM sucrose, 2.5 mM MgCl₂, 100 mM KCl) supplemented with 0.025% w/v digitonin and complete protease inhibitors (Roche) for 10 min on ice to allow permeabilisation of the plasma membrane. Cells were then washed with MELB buffer before fixation in 50 μl eBioscience fixation buffer for 30 min on ice. Cells were washed 1× with eBioscience permeabilization buffer before incubation with an APC‐conjugated anti‐cytochrome‐c antibody (REA702; Miltenyi Biotech) or APC‐conjugated isotype control (REA293; Miltenyi Biotech) for 30 min on ice. Cells were washed 1× before data acquisition on a BD LSR‐Fortessa X20 instrument and analysis using FlowJo.

Speir M., Tye H., Gottschalk T.A., Simpson D.S., Djajawi T.M., Deo P., Ambrose R.L., Conos S.A., Emery J., Abraham G., Pascoe A., Hughes S.A., Weir A., Hawkins E.D., Kong I., Herold M.J., Pearson J.S., Lalaoui N., Naderer T., Vince J.E, & Lawlor K.E. (2023). A1 is induced by pathogen ligands to limit myeloid cell death and NLRP3 inflammasome activation. EMBO Reports, 24(11), e56865.

+ Open protocol

+ Expand

Immunophenotyping of Adipose-Derived Stem Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Surface marker expression was analyzed by flow cytometry. ASCs were used from 4 donors. Cells were incubated at for 4 °C for 15 min in the dark with the following antibodies: CD45 (1:50, REA747), CD73 (1:50, REA804), CD90 (1:50, REA897), CD105 (1:50, REA794) as well as the isotype control (1:50, REA293) (all from Miltenyi Biotec, Germany). Flow cytometry measurement was done on a MACSQuant10 (Miltenyi Biotech) and data was analyzed using FlowLogic (Miltenyi Biotech). A minimum of 10,000 events was analyzed for each sample.

Kaniowska D., Wenk K., Rademacher P., Weiss R., Fabian C., Schulz I., Guthardt M., Lange F., Greiser S., Schmidt M., Braumann U.D., Emmrich F., Koehl U, & Jaimes Y. (2022). Extracellular Vesicles of Mesenchymal Stromal Cells Can be Taken Up by Microglial Cells and Partially Prevent the Stimulation Induced by β-amyloid. Stem Cell Reviews and Reports, 18(3), 1113-1126.

+ Open protocol

+ Expand

Flow Cytometry Staining Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were transferred to FACS tubes (adherent cells were harvested using FACS buffer (PBS pH 7.4 with 0.05% BSA and 2 mM EDTA) prior to staining) with 1 mL of FACS buffer and centrifuged at 150 g for 5 min. Supernatant was decanted, and cells were resuspended in 50 μL of FACS buffer. 10 μL of human IgG (Thermo Fisher 027102) was added, cells were flicked to mix, and were incubated at 4°C for 5 min. Conjugated primary antibody was then added at the manufacturer's recommended dilution, cells were flicked to mix and incubated at 4°C for 30 min. Cells were then washed three times with 1 mL of FACS buffer, centrifuging at 150 g for 5 min and decanting the supernatant after each wash. Cells were resuspended in two drops of FACS buffer prior to flow cytometry. For Miltenyi Biotec antibodies, cells were stained at 4°C for 15 min without blocking and were washed once prior to flow cytometry, as per manufacturer protocol. Antibodies used in this study were as follows: Anti-FLAG-APC (Abcam ab72569), anti-CD2-APC (R&D Systems FAB18561A), anti-CXCR4 (Miltenyi REA649, 130-117-354), anti-CD25-PE (Miltenyi REA945, 130-115-628), anti-SLAM-PE (Miltenyi REA151, 130-123-970), and anti-mouse IgG1-APC (R&D Systems IC002A) or anti-human IgG1-PE (Miltenyi REA293, 130-113-438) were used as isotype controls where appropriate.

Stranford D.M., Simons L.M., Berman K.E., Cheng L., DiBiase B.N., Hung M.E., Lucks J.B., Hultquist J.F, & Leonard J.N. (2024). Genetically encoding multiple functionalities into extracellular vesicles for the targeted delivery of biologics to T cells. Nature biomedical engineering, 8(4).

+ Open protocol

+ Expand

Comprehensive Phenotyping of PCSC Populations

Check if the same lab product or an alternative is used in the 5 most similar protocols

For the determination of ALDH activity, as well as PD-L1/-L2, B2M and HLA-A expressions, the PCSC populations were cultured in growth medium and then partially pre-treated with the cytokine IFNγ (10 ng/mL; Peprotech, Hamburg, Germany) for 72 h. PCSCs were detached via trypsin, harvested, and analyzed via flow cytometry on a Gallios Flow Cytometer (Beckman Coulter Life Sciences, Krefeld, Germany). The following antibodies were used for flow cytometry: anti-CD274 PE (PD-L1; 1:50; MIH1; BD Biosciences, Heidelberg, Germany), anti-CD273 APC-Vio^®770 (PD-L2; 1:10; MIH18; Miltenyi Biotec), anti-B2M PE (1:50; REAL845; Miltenyi Biotec) and anti-HLA-A2 PE (1:50; BB7.2; BD Biosciences), according to manufacturer’s instructions. IsoIgG1-PE (1:10; IS5-21F5; Miltenyi Biotec) and REA Control APC-Vio^®770 (1:10; REA293; Miltenyi Biotec) were used as isotype control antibodies. The measurement of ALDH activity was performed using the ALDEFLUOR™ Kit from STEMCELL Technologies (Vancouver, BC, Canada), in accordance with their guidelines and the approach of Moreb et al. [78 (link)]. For all experiments, dead cells were excluded with Propidium Iodide (PI; 1µg/mL; Thermo Fisher Scientific), and automatic compensation and the resulting analyses were conducted using the evaluation software Kaluza 1.0 from Beckman Coulter Life Sciences.

Witte K.E., Pfitzenmaier J., Storm J., Lütkemeyer M., Wimmer C., Schulten W., Czaniera N., Geisler M., Förster C., Wilkens L., Knabbe C., Mertzlufft F., Kaltschmidt B., am Esch J.S, & Kaltschmidt C. (2021). Analysis of Several Pathways for Efficient Killing of Prostate Cancer Stem Cells: A Central Role of NF-κB RELA. International Journal of Molecular Sciences, 22(16), 8901.

+ Open protocol

+ Expand

Flow Cytometric Analysis of NK Cell Subsets

Check if the same lab product or an alternative is used in the 5 most similar protocols

The cells were stained with the appropriate antibodies and fixed in 1% paraformaldehyde containing phosphate-buffered saline (PBS) at 4 °C for >30 min. Data were obtained using a BD FACSCalibur flow cytometer (BD Biosciences, San Jose, CA, USA) and analyzed using FlowJo version 10 (BD Biosciences). The following antibodies were used: Alexa 488-labeled anti-CD56 (clone B159, BD Pharmingen, Franklin Lakes, NJ, USA), allophycocyanin (APC)-labeled anti-KIR2DL1 (REA284, Miltenyi Biotech), and APC-labeled anti-KIR2DL2/3 (DX27, Miltenyi Biotech, Bergisch, Gladbach, Germany). The isotype control was APC-labeled IgG (REA293, Miltenyi Biotech).

Maeoka R., Nakazawa T., Matsuda R., Morimoto T., Shida Y., Yamada S., Nishimura F., Nakamura M., Nakagawa I., Park Y.S., Tsujimura T, & Nakase H. (2023). Therapeutic Anti-KIR Antibody of 1–7F9 Attenuates the Antitumor Effects of Expanded and Activated Human Primary Natural Killer Cells on In Vitro Glioblastoma-like Cells and Orthotopic Tumors Derived Therefrom. International Journal of Molecular Sciences, 24(18), 14183.

+ Open protocol

+ Expand

Flow Cytometric Characterization of ASCs

Check if the same lab product or an alternative is used in the 5 most similar protocols

Surface marker expression was analyzed by ow cytometry. ASCs were used from 4 donors. Cells were incubated at for 4°C for 15 minutes in the dark with the following antibodies: CD45 (1:50, REA747), CD73 (1:50, REA804), CD90 (1:50, REA897), CD105 (1:50, REA794) as well as the isotype control (1:50, REA293) (all from Miltenyi Biotec, Germany). Flow cytometry measurement was done on a MACSQuant10 (Miltenyi Biotech) and data was analyzed using FlowLogic (Miltenyi Biotech). A minimum of 10,000 events was analyzed for each sample.

Kaniowska D., Wenk K., Rademacher P., Weiß R., Fabian C., Schulz I., Guthardt M., Lange F., Greiser S., Schmidt M., Braumann U., Emmrich F., Koehl U., & Jaimes Y. (2021). Extracellular Vesicles of Mesenchymal Stromal Cells Can be Taken up by Microglial Cells and Partially Prevent The Stimulation Induced by β-Amyloid.

+ Open protocol

+ Expand

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?

Revolutionizing how scientists
search and build protocols!