Easysep mouse pe positive selection kit

Manufactured by STEMCELL

Sourced in United States

The EasySep Mouse PE Positive Selection Kit is a laboratory tool used for the isolation of PE-labeled cells from mouse samples. The kit utilizes magnetic particles and a specialized buffer system to selectively bind and separate the target cells from the rest of the sample.

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Lab products found in correlation

Sca 1 pe antibody, by Thermo Fisher Scientific (2 mentions) Polyacrylamide gel, by Thermo Fisher Scientific (2 mentions) Phosstop phosphatase inhibitor, by Roche (2 mentions) Protease inhibitor, by Roche (2 mentions) Easysep mouse fitc positive selection kit, by STEMCELL (2 mentions) Percoll, by Merck Group (1 mentions) Easysep magnet, by STEMCELL (1 mentions) Anti mouse cd31 antibody, by BioLegend (1 mentions) Anti mouse cd34 antibody, by BioLegend (1 mentions) Easysep mouse apc positive selection kit, by STEMCELL (1 mentions) Anti cd11b microbead, by Miltenyi Biotec (1 mentions) Mojosort mouse cd11c nanobeads, by BioLegend (1 mentions) Papain, by Merck Group (1 mentions) Na pyruvate, by Merck Group (1 mentions) Hepes, by Biosharp (1 mentions) Concanavalin a (cona), by Merck Group (1 mentions) Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions) Lympholyte m, by Cedarlane (1 mentions)

Close spelling variants of this product

EasySep PE Positive Selection Kit Mouse PE Positive Selection kit PE Positive Selection Kit EasySep PE selection kit Mouse PE selection kit EasySep Mouse PE PE selection kit EasySep kits

13 protocols using easysep mouse pe positive selection kit

Purification of Murine Immune Cells

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These lineages were purified from murine BM by positive immuno-magnetic selection:
PE anti-mouse TCR β antibody (clone H57-597) (Biolegend) and EasySep Mouse PE Positive Selection Kit (StemCell Technologies) for T cells, PE anti-mouse CD19 antibody (clone 6D5) (Biolegend) and EasySep Mouse PE Positive Selection Kit (StemCell Technologies) for B cells, anti-CD11b MicroBeads (Miltenyi Biotech) for monocytes, and MojoSort Mouse CD11c Nanobeads (Biolegend) for dendritic cells.

Yu M., Malik Tyagi A., Li J.Y., Adams J., Denning T.L., Weitzmann M.N., Jones R.M, & Pacifici R. (2020). PTH induces bone loss via microbial-dependent expansion of intestinal TNF+ T cells and Th17 cells. Nature Communications, 11, 468.

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Isolation of Liver Cell Subsets

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Hepatic NPCs and hepatocytes were isolated as previously described (Shan et al., 2018 (link)). In brief, mice were anesthetized and liver tissues were perfused with EGTA solution, followed by a 0.04% collagenase digestion buffer. Liver hepatocytes and NPCs were isolated by gradient centrifugation using 35% percoll (Sigma). To further purify LSEC and Mɸs, LSEC and Mɸs fractions were stained with phycoerythrin (PE)-conjugated anti-CD146 (for LSEC, Invitrogen, 12-1469-42), and anti-F4/80 (for Mɸs, Invitrogen, 12-4801-82) antibodies and positively selected using EasySep Mouse PE Positive Selection Kit (Stemcell Technologies) following manufacturer’s instructions. Each subset will yield a purity around 90%.

Shan Z., Li L., Atkins C.L., Wang M., Wen Y., Jeong J., Moreno N.F., Feng D., Gui X., Zhang N., Lee C.G., Elias J.A., Lee W.M., Gao B., Lam F.W., An Z, & Ju C. (2021). Chitinase 3-like-1 contributes to acetaminophen-induced liver injury by promoting hepatic platelet recruitment. eLife, 10, e68571.

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Enrichment and Analysis of AP Cells

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SVF cells were isolated as described above for analysis of phosphorylated AKT, and enrichment for AP cells was achieved using the EasySep Mouse PE Positive Selection Kit (Stem Cell Technologies #18554) using a Sca-1-PE antibody (eBioscience #12-5981-81) according to the manufacturer’s instructions. Positively selected cells were then lysed on ice in 1% IGEPAL with protease inhibitors (Roche) and PhosStop phosphatase inhibitors (Roche). Protein concentration for the resulting lysates was determined using the BCA (bicinchoninic acid) Protein Assay kit from Pierce, and lysates were run on 10% polyacrylamide gels from Invitrogen, followed by western blotting analysis.

Jeffery E., Church C.D., Holtrup B., Colman L, & Rodeheffer M.S. (2015). Rapid Depot-Specific Activation of Adipocyte Precursor Cells at the Onset of Obesity. Nature cell biology, 17(4), 376-385.

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Purification of γδ and αβ T Cells

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WBCs and VAT SVF cells were purified from 24-month old mice (3 pools of 8 aged mice each) and 4-month old mice (3 pools of 10 mice each) for sequential magnetic separation of γδ and αβ T cells. Pools of mice were used to achieve the required starting material for magnetic separation kits. Selection of PE-labeled γδ T cells: γδ T cells were purified by positive selection for γδ TCR with anti-mouse γδ TCR antibody (STEMCELL Technologies 60104PE) and EasySep mouse PE positive selection kit with magnetic particles (STEMCELL Technologies 17666), following the manufacturer’s directions. Magnetically isolated γδ TCR positive cells were washed 5 times to obtain pure cells. Selection of FITC-labeled αβ T cells: The initial supernatant, devoid of γδ T cells and containing the remaining cells, was used to positively select for CD3 positive cells (αβT cells) with anti-mouse CD3 antibody (STEMCELL Technologies 60015FI) and EasySep mouse FITC positive selection kit (STEMCELL Technologies 17668). Magnetically isolated CD3 positive cells were washed 5 times to obtain pure cells. Purity of the fractions was assessed by flow cytometry.

Bruno M.E., Mukherjee S., Powell W.L., Mori S.F., Wallace F.K., Balasuriya B.K., Su L.C., Stromberg A.J., Cohen D.A, & Starr M.E. (2022). Accumulation of γδ T cells in visceral fat with aging promotes chronic inflammation. GeroScience, 44(3), 1761-1778.

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Isolation of Mac1+ Myeloid Cells

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Isolation of Mac1⁺ cells was performed with EasySep Mouse PE Positive Selection Kit (18554, STEMCELL Technologies), EasySep Magnet (18000, STEMCELL Technologies) and PE-conjugated anti-mouse CD11b (M1/70).

Kunimoto H., Meydan C., Nazir A., Whitfield J., Shank K., Rapaport F., Maher R., Pronier E., Meyer S.C., Garrett-Bakelman F.E., Tallman M., Melnick A., Levine R.L, & Shih A.H. (2017). Cooperative Epigenetic Remodeling by TET2 Loss and NRAS Mutation Drives Myeloid Transformation and MEK Inhibitor Sensitivity. Cancer cell, 33(1), 44-59.e8.

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Isolation and transplantation of Vγ4 T cells

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Vγ4 T cells were isolated from lymph node cells and splenocytes of C57BL/6 WT, Il-17a^−/−, and Ifn-γ^−/− mice by the EasySep positive cell isolation systems (PE labeled anti-Vγ4 Ab; BD, USA; EasySep mouse PE positive selection kit; StemCell Technologies, Canada). The purity of isolated Vγ4 T cells was identified by FACS analysis. 1 × 10⁴ cells/wound of WT, Il-17a^−/−, and Ifn-γ^−/− Vγ4 T cells were placed onto the wound bed immediately following excision.

Li Y., Wang Y., Zhou L., Liu M., Liang G., Yan R., Jiang Y., Hao J., Zhang X., Hu X., Huang Y., Wang R., Yin Z., Wu J., Luo G, & He W. (2018). Vγ4 T Cells Inhibit the Pro-healing Functions of Dendritic Epidermal T Cells to Delay Skin Wound Closure Through IL-17A. Frontiers in Immunology, 9, 240.

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Purification of Stromal Vascular Fraction Cell Subsets

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CD31 positive cells from SVF (8–13 aged mice, 2 replicates) were purified by positive selection for CD31 with anti-mouse CD31 antibody (BioLegend 102405) and EasySep mouse FITC positive selection kit (STEMCELL Technologies 17668). Magnetically isolated CD31 positive cells were washed 4 times to obtain pure cells. The initial supernatant containing the remaining cells was then used to positively select for CD34 positive cells with anti-mouse CD34 antibody (BioLegend 119307) and EasySep mouse PE positive selection kit (STEMCELL Technologies 17666). Magnetically isolated CD34 positive cells were washed 3 times to obtain pure cells. The remaining immune cells were purified from the resultant supernatant after CD34 isolation by CD45 antibody and EasySep mouse APC positive selection kit (STEMCELL Technologies, 17667). The resultant supernatant containing non-immune cells was washed 3 times by reapplying to the magnet to obtain pure cells.

Bruno M.E., Mukherjee S., Stromberg A.J., Saito H, & Starr M.E. (2021). Visceral fat-specific regulation of plasminogen activator inhibitor-1 in aged septic mice. Journal of cellular physiology, 237(1), 706-719.

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Isolation and Analysis of Tumor-Infiltrating Immune Cells

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Macrophages or T cells were isolated from 4T1 or EO771 tumors at the experimental end point using EasySep^™ Mouse PE Positive Selection Kit (Stem Cell Technologies, Vancouver, BC) as previously described (29 ). For cell isolation, 1×10⁷–1×10⁸ cells were incubated with 20 μL PE conjugated anti-F4/80 or anti-CD3 (Biolegend) and 50 μL microbeads. The T cells and macrophages were lysed in Qiazol and used for RT-PCR analysis. For ChIP assays, 5–10×10⁶ macrophages were fixed in 1% formaldehyde.

Iwanowycz S., Wang J., Hodge J., Wang Y., Yu F, & Fan D. (2016). Emodin inhibits breast cancer growth by blocking the tumor-promoting feedforward loop between cancer cells and macrophages. Molecular cancer therapeutics, 15(8), 1931-1942.

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Mononuclear Cell Isolation from Mouse Brains

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At peak of disease (approximately day 15 post-immunization), mice were perfused with 10 mL heparinized PBS via direct cardiac puncture and brains isolated. Brains from at least 10 mice were pooled and homogenized using Seward stomacher and mononuclear cells from brains isolated using a 30% percoll (GE Healthcare Life Sciences; Pittsburgh, PA) gradient. Where stated, CD4+ T cells were isolated using Stem Cell Technologies (Cambridge, MA) EasySep™ Mouse PE Positive Selection Kit and phycoerthyrin (PE)-conjugated anti-CD4 (clone: GK 1.5) (purity >90%). Where indicated, total brain mononuclear cells were cultured for 72 h in the presence of 30 μg/mL MOG. Cultures were maintained in complete RPMI 1640 media supplemented with 10% heat-inactivated fetal bovine serum, 10 mM l-glutamine, 10 mM HEPES, 50 μM β-mercaptoethanol, and 100 μg/ml penicillin/streptomycin at 37 °C and 5% CO₂.

Gandy K.A., Zhang J., Nagarkatti P, & Nagarkatti M. (2019). Resveratrol (3, 5, 4′-Trihydroxy-trans-Stilbene) Attenuates a Mouse Model of Multiple Sclerosis by Altering the miR-124/Sphingosine Kinase 1 Axis in Encephalitogenic T Cells in the Brain. Journal of neuroimmune pharmacology : the official journal of the Society on NeuroImmune Pharmacology, 14(3), 462-477.

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Isolation and Enzymatic Analysis of Retinal Rods

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We performed retinal dissociation by incubating dissected retinas in 12 U/ml papain (Sigma, St. Louis MO) and 5 mM L-cysteine in DMEM for 30 minutes on a small vibrating aquarium pump at room temperature. After washing away residual papain with 5 exchanges to fresh DMEM, we triturated the samples with fire-polished Pasteur pipets. We then separated rod photoreceptors with the EasySEP Mouse PE Positive Selection Kit (Stem Cell Technologies, Vancouver Canada) and PE-conjugated anti-CD73 antibodies (Miltenyi Biotec, Bergisch Gladback Germany). We measured the enzymatic activity of NAD⁺-dependent isocitrate dehydrogenase (NAD-IDH/IDH3), alpha-ketoglutarate dehydrogenase (AGDH), and malate dehydrogenase (MDH) in these rods with assay kits from Sigma (St. Louis MO) according to the manufacturer’s instructions. We normalized enzymatic activity to total protein and/or viable rod cell number.

Lin J.B., Kubota S., Ban N., Yoshida M., Santeford A., Sene A., Nakamura R., Zapata N., Kubota M., Tsubota K., Yoshino J., Imai S.I, & Apte R.S. (2016). NAMPT-mediated NAD+ biosynthesis is essential for vision in mice. Cell reports, 17(1), 69-85.

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