Paramagnetic beads

Manufactured by STEMCELL

Sourced in Canada

Paramagnetic beads are small, uniform magnetic particles used in various laboratory applications. They consist of a magnetic core surrounded by a polymer shell. Paramagnetic beads can be easily manipulated using a magnetic field, making them useful for separation, isolation, and purification processes.

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Lab products found in correlation

Anti cd3 cd28, by BD (1 mentions) Lsr 2, by BD (1 mentions) Anti human cd3, by BD (1 mentions) Anti human cd28, by BD (1 mentions) Facsaria, by BD (1 mentions) Anti cd14 pe, by BD (1 mentions) Anti cd19 apc, by BD (1 mentions) Anti cd3 pe, by BD (1 mentions) Facsaria 3 flow cytometer, by BD (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) 96 well cell culture plate, by Corning (1 mentions)

4 protocols using paramagnetic beads

CD4+ T Cell Activation and Autophagy

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CD4 positive T (CD4+ T) cells were isolated from peripheral mononuclear cells using paramagnetic beads (Stemcell technologies) according to the manufacturer’s instructions. Cell purity was > 90%, which was confirmed by BD LSR II (BD Biosciences). CD4⁺ T cells were labeled using carboxyfluorescein succinimidyl ester (CFSE) (BioLegend) according to the manufacturer’s instructions, cultured in the presence of anti-CD3/CD28 (BD Biosciences) (each at final concentration of 2.5 μg/ml) for 2 days and then incubated with A549-atg5-wt or A549-atg5-null cells at a T-cell: A549 cell ratio of 1:5 for the next 2 days. The samples were analyzed by BD LSR II and the results represent the mean ± SD of technical triplicates, each with 30,000 counted cells.

Li R., Zatloukalova P., Muller P., Gil-Mir M., Kote S., Wilkinson S., Kemp A.J., Hernychova L., Wang Y., Ball K.L., Tao K., Hupp T, & Vojtesek B. (2020). The MDM2 ligand Nutlin-3 differentially alters expression of the immune blockade receptors PD-L1 and CD276. Cellular & Molecular Biology Letters, 25, 41.

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CD4+ T Cell Activation and Proliferation

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CD4 positive T cells were obtained from peripheral mononuclear using paramagnetic beads (StemCell Technology, Canada) according to manufacturer's protocol. Cells purity was > 90% confirmed by low Cytometry (LSR II, Becton Dickinson). To determine cell division, CD4⁺ T cells were labeled using Carboxyfluorescein succinimidylester (CFSE, Biolegend, USA) according to manufacturer's instruction. After labeled, T cells were cultured alone in the presence of anti-human CD3 (2.5 μg/ml BD Biosciences) and anti-human CD28 (2.5 μg/ml, BD Biosciences) for 2 days. Before co-culture, SGC-7901 was treated as described in upper functional assay section. Then CD4 positive T cells were collected and incubated with treated SGC-7901 (5:1) for another two days in the presence of anti-human CD3 (2.5 μg/ml, BD Biosciences) and anti-human CD28 (2.5 μg/ml, BD Biosciences). Cell division of CD4⁺ T cells was analyzed by Flow Cytometry (LSR II, Becton Dickinson).

Wang Y., Wang D., Xie G., Yin Y., Zhao E., Tao K, & Li R. (2017). MicroRNA-152 regulates immune response via targeting B7-H1 in gastric carcinoma. Oncotarget, 8(17), 28125-28134.

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Isolation and Characterization of Colon Cancer Immune Cells

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Single cell suspensions were prepared from fresh colon cancer tissues as previously described (Curiel et al., 2004 (link); Curiel et al., 2003 (link); Kryczek et al., 2006 (link); Zou et al., 2001 (link)). Immune cells and tumor cells were enriched using paramagnetic beads (StemCell Technology, Vancouver, Canada). Lin⁻CD45⁻EpCAM⁺ cells primary colon cancer cells and CD4⁺CD45⁺ T cells were sorted from stained single cell suspensions using a high speed cell sorter (FACSaria, Becton Dickinson Immunocytometry Systems, San Jose, CA). Cell purity was >98% as confirmed by flow cytometry (LSR II, BD). Cytokine profile was determined with intracellular staining and analyzed by LSRII (BD).

Kryczek I., Lin Y., Nagarsheth N., Peng D., Zhao L., Zhao E., Vatan L., Szeliga W., Dou Y., Owens S., Zgodzinski W., Majewski M., Wallner G., Fang J., Huang E, & Zou W. (2014). IL-22+CD4+ T cells promote colorectal cancer stemness via STAT3 transcription factor activation and induction of the methyltransferase DOT1L. Immunity, 40(5), 772-784.

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Single-Cell Sorting of HIV VLP-Positive B Cells

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B cell sorting was performed as previously described [18 (link)]. Peripheral blood lymphocytes were isolated by centrifugation on 1.078 density lymphocyte separation medium. CD19⁺ B cells were separated using paramagnetic beads according to the manufacturer’s instructions (STEMCELL Technologies, Vancouver, BC). Then, 2 to 4 x 10⁶ B cells were stained with anti-CD3-PE, anti-CD14-PE, anti-CD19-APC (Becton Dickinson, Franklin Lakes, NJ) and 50 μL of concentrated VLP (containing GFP) preparation on ice for 30 minutes. Flow cytometric analysis and single-cell sorting was performed with a FACSAria III flow cytometer in a Biosafety Level 3 laboratory, equipped with an automated single-cell deposition unit and aerosol containment accessory (Becton Dickinson, Franklin Lakes, NJ). Single HIV VLP⁺/CD19⁺ cells were collected one cell per well into 96-well cell culture plates (Costar^®, Corning Incorporated, Corning, NY), containing RPMI 1640 (Life Technologies, Inc., Rockville, MD) supplemented with 15% gamma-irradiated heat-inactivated fetal bovine serum, 2 mM L-glutamine, 2.5 μg/mL amphotericin B, 60 μg/mL tylosin, 50 μg/mL gentamicin and 0.1% 2-mercaptoethanol.

Hicar M.D., Chen X., Kalams S.A., Sojar H., Landucci G., Forthal D.N., Spearman P, & Crowe JE J.r. (2015). Low frequency of broadly neutralizing HIV antibodies during chronic infection even in quaternary epitope targeting antibodies containing large numbers of somatic mutations. Molecular immunology, 70, 94-103.

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