Cd62l microbeads

Manufactured by Miltenyi Biotec

Sourced in Canada

CD62L microbeads are a type of magnetic bead coated with antibodies specific to the CD62L (L-selectin) receptor. They are used for the isolation and enrichment of CD62L-positive cells from complex samples, such as blood or cell suspensions, through magnetic separation techniques.

Automatically generated - may contain errors

Lab products found in correlation

Cd4 t cell isolation kit 2, by Miltenyi Biotec (2 mentions) Cd4 negative enrichment kits, by STEMCELL (2 mentions) Dynal cd4 t cell negative isolation kit, by Thermo Fisher Scientific (1 mentions) Anti fitc microbeads, by Miltenyi Biotec (1 mentions) Anti cd3 cd28 dynabeads, by Thermo Fisher Scientific (1 mentions) Cd25 microbead kit, by Miltenyi Biotec (1 mentions) Pd l1 ig, by R&D Systems (1 mentions) Celltrace violet, by Thermo Fisher Scientific (1 mentions) Streptavidin microbeads, by Miltenyi Biotec (1 mentions) Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (1 mentions)

7 protocols using cd62l microbeads

Purification and Differentiation of T Cell Subsets

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

CD4⁺ T cells were purified from spleens by magnetic‐activated cell sorting (Dynal CD4⁺ T cell negative isolation kit, Invitrogen, Gent, Belgium) according to the manufacturer's protocol. Naive or memory T cells were purified from previously isolated CD4⁺ T cells subsets by positive or negative selection of CD62L‐expressing cells using the magnetic cell sorting kit (CD62L microbeads, Miltenyi Biotec, Bergisch Gladbach, Germany) according to the manufacturer's protocol. For Treg differentiation experiments, CD25‐positive cells were removed from purified CD4⁺ T cells by magnetic‐activated cell sorting using FITC‐conjugated anti‐CD25 antibodies and anti‐FITC microbeads (Miltenyi Biotec).

Weatherly K., Bettonville M., Torres D., Kohler A., Goriely S, & Braun M.Y. (2015). Functional profile of S100A4‐deficient T cells. Immunity, Inflammation and Disease, 3(4), 431-444.

+ Open protocol

+ Expand

Isolation and Expansion of CD8+ Central Memory T Cells

Cited 1 time

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

CD8⁺CD45RO⁺CD62L⁺ central memory T cells (T_CM) were isolated from peripheral blood mononuclear cells (PBMCs) of healthy donors (Puget Sound Blood Center, Seattle, WA) by negative selection using CD8 isolation kits and CD45RA microbeads and positive selection with CD62L microbeads (Miltenyi Biotec). Following isolation, CD8⁺ T_CM were stimulated with anti CD3/CD28 Dynabeads (Life Technologies) and transduced at a multiplicity of infection (MOI) of 3 on the third day of culture. EGFRt⁺ T_CM subsets were enriched by immunomagnetic selection using biotinylated Erbitux and anti-biotin microbeads (Miltenyi Biotec), then expanded as previously described [44 (link)]. CD8⁺ T_CM were maintained in RPMI medium supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 50 IU/mL recombinant human interleukin 2 (IL-2) and 1 ng/mL recombinant human interleukin 15 (IL-15).

Ravanpay A.C., Gust J., Johnson A.J., Rolczynski L.S., Cecchini M., Chang C.A., Hoglund V.J., Mukherjee R., Vitanza N.A., Orentas R.J, & Jensen M.C. (2019). EGFR806-CAR T cells selectively target a tumor-restricted EGFR epitope in glioblastoma. Oncotarget, 10(66), 7080-7095.

+ Open protocol

+ Expand

T Cell Polarization and Checkpoint Regulation

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

Naïve CD4⁺CD25^negCD62L⁺ T cells were isolated using CD4+ T cell isolation kit II (negative selection), CD25 microbead kit (negative selection), and CD62L microbeads (all from Miltenyi Biotec, Auburn, CA). Isolated CD4⁺CD25^negCD62L⁺ T cells were plated in flasks coated with αCD3 (5 μg/ml) in the presence of αCD28 (5 μg/ml) and Th1-(10 ng/ml IL-12, 1 μg/ml αIL-4), Th2- (10 ng/ml IL-4, μg/ml αIFNγ), or Th17- (20 ng/ml IL-6, 5 ng/ml TGFβ1, 10 ng/ml IL-23, 1 μg/ml αIL-4, 1 μg/ml αIFNγ) skewing cytokines. Cells were cultured for 5 days. Cultures were skewed or stimulated in the presence of either plate bound IgG-Fc (5 μg/ml), or plate bound PD-L1-Ig (R&D Systems, Minneapolis, MN). Cells were rested overnight in the absence of stimulation, and then restimulated on plates coated with various concentrations of αCD3, IgG-Fc and PD-L1-Ig. Cytokine production was assessed by ELISA using eBioscience mAbs.

McAlees J.W., Lajoie S., Dienger K., Sproles A.A., Richgels P.K., Yang Y., Khodoun M., Azuma M., Yagita H., Fulkerson P.C., Wills-Karp M, & Lewkowich I.P. (2015). Differential control of CD4+ T cell subsets by the PD-1/PD-L1 axis in allergic asthma. European journal of immunology, 45(4), 1019-1029.

+ Open protocol

+ Expand

OT-II T Cell Expansion Assay

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

Splenic CD4⁺ T cells derived from OT-II TCR-Tg mice were enriched with Biotin-labeled antibody cocktail (B220, CD8a, CD11b, CD11c, CD19, CD25, NK1.1, and TER-119) with Streptavidin Microbeads (Miltenyi Biotec). CD4⁺ T cells were further incubated with CD62L Microbeads (Miltenyi Biotec) to enrich CD62L⁺CD4⁺ T cells. These CD62L⁺CD4⁺ cells were labeled with 1 μM Cell Trace Violet (Invitrogen) and co-cultured with B cells in the presence of 0.08 and 0.24 μg/ml OVA_323–339-peptide (MBL) for 3 days.

Ono C., Tanaka S., Myouzen K., Iwasaki T., Ueda M., Oda Y., Yamamoto K., Kochi Y, & Baba Y. (2023). Upregulated Fcrl5 disrupts B cell anergy causes autoimmune disease. Frontiers in Immunology, 14, 1276014.

+ Open protocol

+ Expand

Naïve T cell differentiation assay

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

CD4⁺CD62L⁺ naïve T cells were isolated using CD4 negative enrichment kits (Stemcell Technologies, Vancouver, Canada) and CD62L microbeads (Miltenyi Biotec, San Diego, CA) and confirmed to be >95% pure by flow cytometry. These were cultured on 96-well plates pre-coated with anti-CD3 and anti-CD28 under conditions outlined in Supplementary file 2. In particular, the addition of harmine to T_reg^low conditions is abbreviated as T_reg^HAR. Compounds were pinned using a CyBIO CyBi Well Vario (96-well pintool) (Cybio, Jena, Germany). T_reg and Th1 cultures were fed with equal volume of IL-2-supplemented media (10 ng/ml) and retreated with compound at day 2, split 1:2 into IL-2-supplemented media at day 3 and analyzed at day 4. Th17 and Th0 cultures were treated similarly except no IL-2 was supplemented. Cell proliferation was monitored using CFSE (Life Technologies, Carlsbad, CA) per manufacturer's instructions.

Khor B., Gagnon J.D., Goel G., Roche M.I., Conway K.L., Tran K., Aldrich L.N., Sundberg T.B., Paterson A.M., Mordecai S., Dombkowski D., Schirmer M., Tan P.H., Bhan A.K., Roychoudhuri R., Restifo N.P., O'Shea J.J., Medoff B.D., Shamji A.F., Schreiber S.L., Sharpe A.H., Shaw S.Y, & Xavier R.J. (2015). The kinase DYRK1A reciprocally regulates the differentiation of Th17 and regulatory T cells. eLife, 4, e05920.

+ Open protocol

+ Expand

Isolation and Characterization of Antigen-Specific Memory CD4+ T Cells

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

To analyze mouse primary CD4⁺ T cells, naive (CD62L^hiCD44^lo) or memory (CD62L^loCD44^hiCD25⁻) phenotype CD4⁺ T cells were prepared from spleen via MACS (CD4⁺ T cell isolation kit II and CD62L microbeads [Miltenyi]) and/or flow cytometric cell sorting. To generate OVA-specific memory CD4⁺ T cells, CD4⁺ T cells from OVA-specific OT-II TCR Tg mice were utilized. Naive WT or Trp53^−/− OT-II CD4⁺ T cells (CD45.2) were purified and 5 × 10⁵ cells were adoptively transferred (i.v.) to CD45.1 or Rag1^−/− mice. Recipient mice were immunized with OVA-alum (i.p.) 1 day after cell transfer. Three to four weeks later, CD44^hiCD62L^loCD25⁻ memory phenotype CD4⁺ T cells (CD45.2) were isolated by MACS from splenocytes of the immunized mice.

Watanabe M., Moon K.D., Vacchio M.S., Hathcock K.S, & Hodes R.J. (2014). Downmodulation of Tumor Suppressor p53 by T Cell Receptor Signaling Is Critical for Antigen-Specific CD4+ T Cell Responses. Immunity, 40(5), 681-691.

+ Open protocol

+ Expand

Autophagy-Regulated Naive T Cell Activation

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

CD4⁺ CD62L⁺ naïve T cells were isolated from C57BL/6J (WT), Lck Cre⁺x Atg16L1^flox/flox (cKO), and Atg16L1^T300A/T300A mutant (T300A) mice using CD4 negative enrichment kits (Stemcell Technologies, Vancouver, Canada) and CD62L microbeads (Miltenyi Biotec, San Diego, CA). For each genotype, the cells were prepared independently from 3 mice as biological replicates. T cells isolated per genotype each received no stimulation, CD3/CD28 dynabeads (for 4h or 20h) or 1 μM Torin treatment (for 4h). Peak T cell activation, as defined by number of differentially expressed genes, after CD3/CD28 stimulation was observed at 20h; the 4h time point allowed us to perform a time-varying analysis of transcriptional programs in response to CD3/CD28 stimulation. A 4h Torin treatment activated autophagy in T cells.

Varma M., Kadoki M., Lefkovith A., Conway K.L., Gao K., Mohanan V., Tusi B.K., Graham D.B., Latorre I.J., Tolonen A.C., Khor B., Ng A.C, & Xavier R.J. (2020). Cell type- and stimulation-dependent transcriptional programs regulated by Atg16L1 and its Crohn’s disease risk-variant T300A. Journal of immunology (Baltimore, Md. : 1950), 205(2), 414-424.

+ 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!