Single-cell suspensions were prepared from the spleens of donor CD45.1+ or CD45.2+ TCRβ-transgenic EF4.1 mice or CD45.2+ Nur77-GFP EF4.1 doubly transgenic mice, and CD4+ T cells were enriched using an immunomagnetic positive selection kit (StemCell Technologies, Vancouver, BC, Canada), at >90% purity. Donor transgenic CD4+ T cells (1 × 106 per recipient) were injected into recipient mice intravenously.
Immunomagnetic positive selection kit
Manufactured by STEMCELL
Sourced in Canada
The Immunomagnetic positive selection kit is a laboratory tool used to isolate and purify specific cell types from complex samples. It utilizes magnetic beads coated with antibodies that bind to target cell surface markers, allowing for the separation and enrichment of the desired cell population.
Automatically generated - may contain errors
Lab products found in correlation
Immunomagnetic enrichment kit, by STEMCELL (2 mentions)
Rpmi 1640, by GE Healthcare (2 mentions)
Methylcellulose medium, by STEMCELL (1 mentions)
Methocult h4434 classic, by STEMCELL (1 mentions)
Interleukin 3 (il 3), by Thermo Fisher Scientific (1 mentions)
Ficoll density gradient centrifugation, by GE Healthcare (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Interleukin 6 (il 6), by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Stemspan sfem, by STEMCELL (1 mentions)
6 protocols using immunomagnetic positive selection kit
1
Adoptive Transfer of Transgenic CD4+ T Cells
2
Clonogenic Assay of miR-375 in AML
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Bone marrow mononuclear cells (blasts% > 70%) from six AML patients were harvested. CD34+ cells were enriched by immunomagnetic positive selection kit (Stem Cell Technologies), followed by the retrovirus transduction of MSCV-miR-375 or MSCV-NC. Retrovirus-transduced CD34+ cells (5×103) were seeded on a 35-mm dish with 1.0 ml of methylcellulose medium (MethoCult™ H4434 Classic, Stem Cell Technologies). HL-60 and THP1 cells, which were transduced with MSCV-miR-375 or MSCV-NC, were seeded in the same methylcellulose medium (Stem Cell Technologies). Normal CD34+ cells were isolated and enriched from UCB, followed by the retrovirus transduction of MSCV-miR-375 or MSCV-NC and then were plated in the same methylcellulose medium (Stem Cell Technologies). Colonies were scored 10–14 days after plating according to the manufacturer’s protocol.
3
Exploring Leukaemic Cell Lines and AML Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human leukaemic cell lines THP1 and MV4‐11 (ATCC, Manassas, VA, USA) were used for the present study. All leukaemic cell lines were cultured in a humidified 37°C incubator with 5% CO2 in RPMI 1640 (Invitrogen, Carlsbad, CA, USA) with 10% fetal bovine serum (Sigma‐Aldrich, St. Louis, MO, USA). Bone marrow (BM) mononuclear cells from AML patients were isolated by Ficoll density gradient centrifugation (GE Healthcare, Uppsala, Sweden) and cultured in StemSpan SFEM (Stemcell Technologies, Vancouver, Canada) supplemented with human recombinant interleukin‐3 (IL‐3, PeproTech, Rocky Hill, NJ, USA), stem cell factor (SCF, PeproTech), and interleukin‐6 (IL‐6, PeproTech) at final concentrations of 10 ng/mL. All the patients gave informed consent. Normal human CD34+ HSPCs were isolated from umbilical cord blood (UCB) and enriched by an immunomagnetic positive selection kit (Stemcell Technologies). Bort (MCE, Princeton, NJ, USA), Palbociclib (MCE), and Bay 11‐7082 (MCE) were dissolved in dimethyl sulfoxide (DMSO) and kept at −20°C until used. All procedures in our studies involving human participants were following the Ethics Committee of the First Affiliated Hospital of Wenzhou Medical University and the Declaration of Helsinki. The clinical characteristics of AML patients are summarized in Table S1 .
4
Adoptive Transfer of Transgenic CD4+ T Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Single-cell suspensions were prepared from the spleens of donor CD45.1+ or CD45.2+ TCRβ-transgenic EF4.1 mice and CD4+ T cells were enriched using an immunomagnetic positive selection kit (StemCell Technologies, Vancouver, Canada), at >90% purity. Donor transgenic CD4+ T cells (1×106 per recipient) were injected into recipient mice intravenously.
5
Enrichment of Murine Hematopoietic Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Bone marrow was flushed from femur and tibia, and suspended in phosphate buffered saline. RBCs were removed by ammonium chloride-lysis solution. Lineage-negative (Lin−) population of cells was isolated by negative selection by using immunomagnetic enrichment kit (StemCell Technologies) as per the manufacturer’s instructions. Briefly, the mononuclear cells were suspended in the recommended medium and incubated with specific antibodies for labeling unwanted nonhematopoietic stem/progenitor cells which include CD5, CD11b, CD19, CD45R, 7–4, Ly-6G/C (Gr-1), and TER119-expressing cells. Antibody-treated cells were then labeled by Tetrameric Antibody Complexes that recognize biotin and dextran-coated magnetic particles. Unwanted labeled cells were then separated by using an EasySep™ magnet. Thus the desired Lin− cells were obtained by negative selection. These cells were then enriched for cKit+ cells by using positive immunomagnetic selection kit (StemCell Technologies). LK cells were plated in RPMI1640 (GE Healthcare) in U-bottom, 96-well plate at a low density of 2 × 104 cells/150 μL per well, until they are used for proliferation or migration assay in less than 24 hours following isolation. Representative flow cytometric dot plots evaluating the purity of LK cells enriched by this method is shown in Fig. 9 .
6
Isolating Lin− Sca-1+ cKit+ Hematopoietic Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Lin− cells were isolated by negative selection by using the immunomagnetic enrichment kit (StemCell Technologies Inc.). Briefly, bone marrow monocyte cell suspension was prepared in recommended medium at required dilution and incubated with antibodies for binding CD5-, CD11b-, CD19-, CD45R/B220-, Ly-6G/C (Gr-1)-, and TER119-expressing cells. These cells are then labeled by tetrameric antibody complexes that recognize biotin and dextran-coated magnetic particles, and antibody-bound cells were then separated by using EasySep™ magnet, thus obtaining Lin− cells. Lin− cells were then enriched for Sca-1+ and cKit+ cells by using positive immunomagnetic selection kit (StemCell Technologies) as reported before [33 (link)]. Cells were then plated in RPMI1640 (GE Healthcare) in U-bottom, 96-well plate at a low density of 2 × 104 cells/150 μL per well, until they are used for proliferation or migration assay for less than 24 h following isolation.
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!