Moflo xdp

Manufactured by BD

Sourced in United States

The MoFlo XDP is a high-performance cell sorter designed for advanced flow cytometry applications. It features a flexible and modular design to accommodate a wide range of research needs. The MoFlo XDP provides precise cell sorting capabilities with high purity and recovery rates.

Automatically generated - may contain errors

Lab products found in correlation

Ter119 ter 119 apccy7, by BD (2 mentions) Fusion cytometer, by BD (2 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) Lsrii flow cytometer, by BD (1 mentions) Fortessa analyser, by BD (1 mentions) Lineage cell depletion kit, by Miltenyi Biotec (1 mentions) Fortessa, by BD (1 mentions) Immunoseq platform, by Adaptive Biotechnologies (1 mentions) Immunoseq assay, by Adaptive Biotechnologies (1 mentions) Dneasy blood and tissue kit, by Qiagen (1 mentions) Fixable fluorescent viability stain, by Thermo Fisher Scientific (1 mentions) Influx cytometer, by BD (1 mentions) Facsariatm 2, by BD (1 mentions) Pmd2 g plasmid, by Addgene (1 mentions) Lumina in vivo imaging system, by PerkinElmer (1 mentions) Polyethyleneimine, by Polysciences (1 mentions) Pspax2, by Addgene (1 mentions) Sybr green jumpstarttm taq readymixtm, by Merck Group (1 mentions) Stepone plus pcr, by Thermo Fisher Scientific (1 mentions)

18 protocols using moflo xdp

Purification of Naïve and Treg CD4+ T Cells

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Bulk CD4⁺ T cells were purified from the spleen and mLN by negative selection as previously described (Coccia et al., 2012 (link)). Naïve CD4⁺ T cells were then sorted as CD4⁺ CD25^- CD44^- CD62L⁺. T_reg cells were sorted as CD4⁺ CD25⁺ when sorted from Atg16l1^ΔCD4 and Atg16l1^fl/fl mice and as CD4⁺ YFP⁺ when sorted from Atg16l1^ΔFoxp3 and Foxp3^Cre mice. Cells were sorted using an Astrios, Beckman Coulter MoFlo XDP or AriaIII BD Bioscience. Post-sort flow cytometry analyses confirmed that the purity of sorted populations was >97%.

Kabat A.M., Harrison O.J., Riffelmacher T., Moghaddam A.E., Pearson C.F., Laing A., Abeler-Dörner L., Forman S.P., Grencis R.K., Sattentau Q., Simon A.K., Pott J, & Maloy K.J. (2016). The autophagy gene Atg16l1 differentially regulates Treg and TH2 cells to control intestinal inflammation. eLife, 5, e12444.

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Intracellular Arsenic Measurement

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K562 cells were incubated with or without ee-As₄S₄ for different time. After incubation, viable single cells were collected by a MoFlo XDP flow sorter (BD Biosciences, San Jose, CA, USA). The arsenic content in the collected cells was measured using hydride generation atomic fluorescence spectrometry (AFS-8230 HG-AFS, Beijing Titan Instrument Co. Ltd., Beijing, China).

Wang T., Wen T., Li H., Han B., Hao S., Wang C., Ma Q., Meng J., Liu J, & Xu H. (2019). Arsenic sulfide nanoformulation induces erythroid differentiation in chronic myeloid leukemia cells through degradation of BCR-ABL. International Journal of Nanomedicine, 14, 5581-5594.

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Engineered Jurkat Cell Lines for Immunotherapy

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Jurkat cells (acute T cell leukaemic cell line from ATCC) were modified to express CD3⁺CD7⁺ CD3⁺CD7⁻, CD3⁻CD7⁺ and CD3⁻CD7^– by SpCas9 mRNA-based disruption of TCR and/or CD7 using TRBC sgRNA and CD7 sgRNA and maintained in culture in RPMI-1640 (Thermo Fisher Scientific) supplemented with 10% FBS (Millipore Sigma) (Supplementary Fig. 9). For in vivo experiments cells were stably transduced with a 3rd generation pCCL-PGK-EGFP-LUC lentiviral vector and were sorted on a MoFlo XDP (BD) for GFP expression prior to banking.

Georgiadis C., Rasaiyaah J., Gkazi S.A., Preece R., Etuk A., Christi A, & Qasim W. (2021). Base-edited CAR T cells for combinational therapy against T cell malignancies. Leukemia, 35(12), 3466-3481.

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Flow Cytometry Analysis of SC-β and ANGI Cells

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Flow cytometry analysis of differentiation efficiencies was performed using the LSRII flow cytometer by BD. FACS purification of pre- and post-transplant SC-β cells was performed using a combination of MoFlo Astrios, MoFlo XDP, and BD FASCARIA III sorting instruments. Cells were first gated on side and forward scatter profiles followed by gating on GFP-positive cells and subsequent TSQ (405 nm)-positive PI-negative populations following the protocol (Davis et al., 2019 (link)). Cells were sorted into stage 6 medium and centrifuged/washed before lysis in TriZOL RNA reagent for extraction using trizol/chloroform extraction.
Flow cytometry for ANGIs differentiations were performed using the Attune NXT and sorted using the above-mentioned instruments. Gating strategies for ANGIs analysis was to first plot mCherry vs violet or empty channel to identify single positive cells and the mCherry⁺ subset was then plotted for eGFP vs empty channel or violet. Flow cytometry staining for ANGIs used the above antibodies as well as antibodies against endocrine hormones SST, PPY, GHRL in the far-red channel. These experiments included antibodies to mCherry and eGFP to enhance signal after fixation.

Davis J.C., Ryaboshapkina M., Kenty J.H., Eser P.Ö., Menon S., Tyrberg B, & Melton D.A. (2024). IAPP Marks Mono-hormonal Stem-cell Derived β Cells that Maintain Stable Insulin Production in vitro and in vivo. bioRxiv.

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Multicolor Flow Cytometry Analysis

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Flow cytometry was performed using a BD Fortessa analyser with subsequent data analysis using FlowJo 7.6.5 software. Cell sorting was performed using a MoFlo XDP (BD) cell sorter. mCherry, Green fluorescent protein (GFP) and blue fluorescent protein (BFP) were excited using 561 nm, 488 nm and 405 nm laser and detected using a 610/20, 530/30 and 440/40 filter.

Gao X., Tsang J.C., Gaba F., Wu D., Lu L, & Liu P. (2014). Comparison of TALE designer transcription factors and the CRISPR/dCas9 in regulation of gene expression by targeting enhancers. Nucleic Acids Research, 42(20), e155.

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Murine Malaria Infection Monitoring

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Malaria: Mice (male) housed under reverse light were infected intraperitoneally with 10⁵Plasmodim chabaudi chabaudi AS infected red blood cells (RBC). Blood smears were taken daily (am) when parasite sequestration is low in order to best estimate the level of parasitemia, the proportion of parasitized RBCs were monitored as described previously51 (link). Measurements of clinical pathology, body weight and temperature (measured using a non-invasive thermometer, Fluke), were also taken daily (pm) and calculated relative to the day 0 baseline. For RNA-seq and ATAC-seq, T cells were enriched by negative selection from individual spleens (3 mice each Maf^fl/fl and Maf^fl/flCd4-cre) on day 7 post infection as described above and Ter119-CD3⁺CD4⁺ T cells (Ter119 (TER-119) APCCy7, BD; CD3 (145-2C11) APC, CD4 (RM4-5) e450, eBioscience) were sorted on MoFlo™ XDP or BD Fusion cytometer.

Gabryšová L., Alvarez-Martinez M., Luisier R., Cox L.S., Sodenkamp J., Hosking C., Pérez-Mazliah D., Whicher C., Kannan Y., Potempa K., Wu X., Bhaw L., Wende H., Sieweke M.H., Elgar G., Wilson M., Briscoe J., Metzis V., Langhorne J., Luscombe N.M, & O’Garra A. (2018). c-Maf controls immune responses by regulating disease-specific gene networks and repressing IL-2 in CD4+ T cells. Nature immunology, 19(5), 497-507.

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Immunophenotyping mouse hematopoietic cells

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Single-cell suspensions were incubated with purified anti-CD16/32 (clone 93) for 10 min on ice to block Fc receptors. Fluorochrome- or biotin-labeled monoclonal antibodies (clones denoted in parentheses) against B220 (RA3-6B2), CD19 (1D3), CD3ɛ (145-2C11), CD4 (RM4-5), CD8 α (53–6.7), TCRβ (B20.6), NK1.1 (PK136), CD11b (M1/70), Gr1 (RB6-8C5), Ter119 (TER-119), Sca1 (D7), c-kit (2B8), CD48 (HM48-1), CD150 (TC15-12 F 12.2), CD34 (RAM34), and Flt3 (A2F10) were purchased from BD Biosciences, BioLegend or eBioscience. Cells were stained with antibodies on ice for 20 min before washing. Cells were run on a Fortessa (BD Biosciences) or MoFlo XDP (BD Biosciences) and analyzed by Flowjo (Tree Star). For HSC sorting, Lin⁻ cells were enriched by Lineage Cell Depletion Kit (Miltenyi Biotec) before antibody staining.

Tsang J.C., Yu Y., Burke S., Buettner F., Wang C., Kolodziejczyk A.A., Teichmann S.A., Lu L, & Liu P. (2015). Single-cell transcriptomic reconstruction reveals cell cycle and multi-lineage differentiation defects in Bcl11a-deficient hematopoietic stem cells. Genome Biology, 16, 178.

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Murine Malaria Infection Monitoring

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GFP-Driven Cell Sorting and Analysis

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GFP was used as a reporter gene for transfected cells, and positive cells were sorted using a Beckman Coulter MoFlo XDP or a BD FACSVantage Diva system. The sorted cells were used for RT-qPCR, Western blotting, and cell-cycle analysis [propidium iodide (PI)/BrdU] as described in Supporting Information.

Ezerskyte M., Paredes J.A., Malvezzi S., Burns J.A., Margison G.P., Olsson M., Scicchitano D.A, & Dreij K. (2018). O6-methylguanine–induced transcriptional mutagenesis reduces p53 tumor-suppressor function. Proceedings of the National Academy of Sciences of the United States of America, 115(18), 4731-4736.

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Profiling Antigen-Specific CD8+ T cells

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Antigen-specific CD8⁺ T cells were sorted based on MART-1 tetramer expression using the MoFlo XDP (BD Biosciences). Total genomic DNA was isolated with the DNeasy Blood and Tissue Kit (Qiagen, Venlo, the Netherlands). The DNA was used for deep sequencing of the β-chain, using the ImmunoSEQ platform (Adaptive Biotechnologies, Seattle, WA, USA). Details of the deep sequencing assay are as follows: the TCRβ and TCRγ CD3 region was amplified and sequenced using the ImmunoSEQ assay (Adaptive Biotechnologies, Seattle, WA). Specifically, CD8⁺ T cells from patients were either sequenced pre-expansion or expanded for MART-1 antigen using DCs or nano-aAPC, before sorting for MART-1 tetramer positive cells, and then sequenced. Diversity metrics, clone distribution, and Vβ gene usages were examined both pre- and post-expansion for each patient. The diversity metrics that we examined were: productive entropy, which is Shannon’s Entropy (

H = - \sum p_{i} {log}_{2} p_{i}

where p_i is the frequency of a given clone) for all productive rearrangements within a sample, with larger numbers representing greater diversity; and productive clonality (

C = 1 - \frac{H}{{log}_{2} N}

where H is productive entropy and N is the number of unique CDR3 clones) which ranges from 0 to 1 with lower values representing greater diversity.

Ichikawa J., Yoshida T., Isser A.Y., Laino A.S., Vassallo M., Woods D.M., Kim S., Oelke M., Jones K., Schneck J.P, & Weber J.S. (2020). Rapid Expansion of Highly Functional Antigen-Specific T cells from Melanoma Patients by Nanoscale Artificial Antigen Presenting Cells. Clinical cancer research : an official journal of the American Association for Cancer Research, 26(13), 3384-3396.

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