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Biotin selection kit

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The Biotin Selection Kit is a laboratory tool designed for the purification and enrichment of biotinylated cells or molecules. It allows for the efficient isolation and separation of biotin-labeled target cells or proteins from a complex mixture. The kit contains essential components for the magnetic separation process, enabling researchers to obtain highly purified samples for further analysis and experimentation.

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Lab products found in correlation

Murine egf, by Thermo Fisher Scientific (2 mentions) Biotinylated anti mouse gr1, by STEMCELL (2 mentions) B27 supplement, by Thermo Fisher Scientific (2 mentions) Murine bfgf, by Thermo Fisher Scientific (2 mentions) Mouse cd4 t cell isolation kit, by STEMCELL (1 mentions) Gm csf, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

EasySep Biotin Selection Kit (20 citations) EasySep Biotin Positive Selection Kit (14 citations) EasySep Mouse Biotin Positive Selection Kit (8 citations) EasySep Mouse Biotin Positive Selection Kit II (5 citations) EasySep mouse biotin selection kit (5 citations) EasySep Release Human Biotin Positive Selection Kit (4 citations) EasySep Human Biotin Positive Selection Kit II (3 citations) Mouse Biotin Selection Kit (2 citations) Biotin based selection kit (2 citations) EasySep Biotin Positive Selection Kit II (2 citations)

5 protocols using biotin selection kit

1

Tumor Cell-MDSC Co-Culture Protocol

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Tumor cells (20,000 cells per well) were cultured in low-adhesion 24well plate in DMEM/F12 medium with murine bFGF (20 ng mL−1, Life Technologies), murine EGF (20 ng mL−1, Life Technologies) and B27 supplement (GIBCO). For co-culture, 80,000 Gr1+ MDSCs were added. As indicated, in some instances MDSCs and tumor cells were co-cultured in Boyden chambers, putting MDSCs on insets with 0.4 μm pore size membranes. Co-culture periods were 5 h for RNA expression study, 24 h for FACS of cancer stem cell surface markers, and 6 to 12 d to determine mammosphere forming units. As indicated in figures, specific control groups received Gr1+ cells of TF mice (NN, normal neutrophils) instead of Gr1+ MDSCs. These control cells were isolated from bone marrow with the same magnetic sorting procedure used for MDSCs (Biotinylated anti-mouse Gr1, BD Pharmingen, Biotin Selection Kit, Stem Cell Technologies). Mammospheres were enumerated by manual counting of low magnification images of the cultures (Cellcount). The images in Supplementary Fig. 7a are representative of over 20 images obtained from different experiments. For flow cytometry, mammospheres were dissociated by trypsin-EDTA digestion. Total tumor cell counts were obtained by FACS of the GFP-tagged tumor cells.
Welte T., Kim I.S., Tian L., Gao X., Wang H., Li J., Holdman X.B., Herschkowitz J.I., Pond A., Xie G., Kurley S., Nguyen T., Liao L., Dobrolecki L.E., Mo Q., Edwards D.P., Huang S., Xin L., Xu J., Li Y., Lewis M.T., Wang T., Westbrook T.F., Rosen J.M, & Zhang X.H. (2016). Oncogenic mTOR signaling recruits myeloid-derived suppressor cells to promote tumor initiation. Nature cell biology, 18(6), 632-644.
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2

Isolation and co-culture of CD4+ T cells and BMDCs

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For the isolation of CD4+ T cells from cLN, cLNs were harvested from ligature mice (day 14) and the mouse CD4+ T cell Isolation Kit (STEMCELL Technologies) was used to isolate cells from single cell suspensions prepared from the cLNs. For the preparation of the bone marrow–derived dendritic cells (BMDMs), murine bone marrow cells were cultured in the RPMI/10% FBS containing recombinant murine 10 ng/mL GM-CSF (PeproTech, Rocky Hill, NJ) for 6 days. Loosely attached DC cells were then collected using the biotin selection kit (STEMCELL Technologies) and enriched with biotinylated CD11c monoclonal antibody (Thermo Fisher Scientific, Waltham, MA). BMDCs were pulsed with heat-denatured bacterial antigens for 18 h at 37°C. Antigen-pulsed BMDCs and cLN-derived CD4+T cells were co-cultured for 24 h at 37°C. Cytokines in supernatants were quantified by ELISA.
Kitamoto S., Nagao-Kitamoto H., Jiao Y., Gillilland MG I.I.I., Hayashi A., Imai J., Sugihara K., Miyoshi M., Brazil J.C., Kuffa P., Hill B.D., Rizvi S.M., Wen F., Bishu S., Inohara N., Eaton K.A., Nusrat A., Lei Y.L., Giannobile W.V, & Kamada N. (2020). The Intermucosal Connection between the Mouth and Gut in Commensal Pathobiont-Driven Colitis. Cell, 182(2), 447-462.e14.
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3

Pmel T Cell Proliferation Assay

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Splenic cells of Pmel TCR transgenic mice were collected and CD8+ Pmel T cells were enriched by negative CD8 T cell enrichment kit (Stemcell technologies). Naïve CD8+ Pmel T cells were further purified by removing CD44+ cells with biotin selection kit (Stemcell technologies). Purified naïve Pmel cells were then loaded with 2 M CFSE at room temperature for 15 mins. CFSE-labeled naïve CD8+ Pmel T cells were mixed with tumor-infiltrating DCs in a 5:1 ratio in the presence of 1μg/ml gp100. For agonistic anti-CD40 antibody, the cell mixtures were incubated with 50μg/ml of IgG (Bioxcell) or agonistic anti-CD40 antibody (FGK45) (Bioxcell) during this period. Proliferation of CD8+ Pmel T cells was determined by FACS analysis to measure CFSE dilution.
Ho P.C., Meeth K.M., Tsui Y.C., Srivastava B., Bosenberg M.W, & Kaech S.M. (2014). Immune-based antitumor effects of BRAF inhibitors rely on signaling by CD40L and IFNγ. Cancer research, 74(12), 3205-3217.
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4

Isolation of Brain-Reactive Memory B Cells

Cited 1 time
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Isolation of single human memory antigenic-specific B cells was performed as previously described22 (link) with several modifications. B cells were purified from fresh mononuclear cells by negative selection using a B cell kit (StemCell Technology, Vancouver, BC, Canada). They were then incubated for 30 min at room temperature with human fetal brain lysate (3 μg ml−1, Nouvus, Littleton, CO, USA) labeled with biotin using the EZ-Link Sulfo-NHS-Biotin labeling kit (Life Technologies, Carlsbad, CA, USA). Cells bound by biotinylated brain antigens were isolated with a biotin selection kit (StemCell Technologies) and stained with fluoroscein isothiocyanate-conjugated anti-human CD19, phycoerythrin-conjugated anti-human CD27 and allophycocyanin streptavidin to allow the separation of CD19+, CD27+, brain lysate+ memory B cells. As a control, the fraction that initially was identified as nonbrain-reactive was incubated with biotinylated brain antigen and stained as described above. No allophycocyanin-positive cells were detected in this fraction. Finally, CD19+, CD27+, allophycocyanin+ single cells were isolated on a BD FACSAria as described in ref. 23 (link).
Brimberg L., Mader S., Jeganathan V., Berlin R., Coleman T., Gregersen P., Huerta P., Volpe B, & Diamond B. (2016). Caspr2-reactive antibody cloned from a mother of an ASD child mediates an ASD-like phenotype in mice. Molecular psychiatry, 21(12), 1663-1671.
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5

Tumor Cell-MDSC Co-Culture Protocol

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Tumor cells (20,000 cells per well) were cultured in low-adhesion 24well plate in DMEM/F12 medium with murine bFGF (20 ng mL−1, Life Technologies), murine EGF (20 ng mL−1, Life Technologies) and B27 supplement (GIBCO). For co-culture, 80,000 Gr1+ MDSCs were added. As indicated, in some instances MDSCs and tumor cells were co-cultured in Boyden chambers, putting MDSCs on insets with 0.4 μm pore size membranes. Co-culture periods were 5 h for RNA expression study, 24 h for FACS of cancer stem cell surface markers, and 6 to 12 d to determine mammosphere forming units. As indicated in figures, specific control groups received Gr1+ cells of TF mice (NN, normal neutrophils) instead of Gr1+ MDSCs. These control cells were isolated from bone marrow with the same magnetic sorting procedure used for MDSCs (Biotinylated anti-mouse Gr1, BD Pharmingen, Biotin Selection Kit, Stem Cell Technologies). Mammospheres were enumerated by manual counting of low magnification images of the cultures (Cellcount). The images in Supplementary Fig. 7a are representative of over 20 images obtained from different experiments. For flow cytometry, mammospheres were dissociated by trypsin-EDTA digestion. Total tumor cell counts were obtained by FACS of the GFP-tagged tumor cells.
Welte T., Kim I.S., Tian L., Gao X., Wang H., Li J., Holdman X.B., Herschkowitz J.I., Pond A., Xie G., Kurley S., Nguyen T., Liao L., Dobrolecki L.E., Mo Q., Edwards D.P., Huang S., Xin L., Xu J., Li Y., Lewis M.T., Wang T., Westbrook T.F., Rosen J.M, & Zhang X.H. (2016). Oncogenic mTOR signaling recruits myeloid-derived suppressor cells to promote tumor initiation. Nature cell biology, 18(6), 632-644.
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