iPS cells were maintained and differentiated as previously described (see Supplementary data ) 37 (link). At day 18 neural precursor cells (NPCs) were isolated using Anti-PSA-NCAM MicroBeads (30-092-966¸ Miltenyi Biotec) according to the manufacturer’s protocol. Sorted cells were fluorescently stained with an Anti-IgM antibody conjugated with PE dye (130-095-908; Miltenyi Biotec) and analysed by flow cytometry using a GUAVA EasyCyte 6-2L instrument to assess purity of the recovered population. Unlabelled cells were used as negative control. Cultures were harvested at day 70 for analysis of mature neurons. To obtain a more pure population of neurons for qRT-PCR, mature cultures were sorted using a CD24 MicroBead Kit (130-095-951, Miltenyi Biotec)39 (link). Sorted neurons were plated on poly-ornithine and laminin coated slides and allowed to attach for 48 hours before being stained for β-III-Tubulin (Chemicon) to confirm their identity.
Cd24 microbead kit
Manufactured by Miltenyi Biotec
Sourced in Germany
The CD24 Microbead Kit is a laboratory equipment product designed for the isolation and enrichment of CD24-positive cells from various sample types. The product utilizes magnetic beads coated with antibodies specific to the CD24 surface antigen. The core function of this kit is to enable the efficient separation and purification of CD24-expressing cells from a heterogeneous cell population.
Automatically generated - may contain errors
Lab products found in correlation
Cd44 microbeads, by Miltenyi Biotec (4 mentions)
Minimacs columns, by Miltenyi Biotec (2 mentions)
Goat anti mouse igg microbeads, by Miltenyi Biotec (2 mentions)
Anti psa ncam microbeads, by Miltenyi Biotec (1 mentions)
β tubulin 3, by Merck Group (1 mentions)
Quadromacs separator, by Miltenyi Biotec (1 mentions)
Anti pe micro bead, by Miltenyi Biotec (1 mentions)
Anti cd44 pe, by BioLegend (1 mentions)
Macs separator, by Miltenyi Biotec (1 mentions)
Macs column, by Miltenyi Biotec (1 mentions)
70 m cell strainer, by BD (1 mentions)
8 protocols using cd24 microbead kit
1
Efficient Isolation and Characterization of Mature Neurons
2
Isolation of Breast Cancer Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
1 × 107 cells from 4T1 spheres, as well as monolayer culture and MDAMB-231 spheres, were incubated with primary antibody against CD24 according to the manufacturer's procedure mentioned in CD24 Microbead Kit (MiltenyiBiotec). Following that, goat anti-mouse IgG microBeads (Miltenyi Biotec) were added to the labeled cells and MiniMACS columns (Miltenyi Biotec) were used to magnetically separate the cells. CD44 microbeads (Miltenyi Biotec) were added to acquire CD24− cells at 4 °C for 15 min. Cells were again washed and magnetically separated. The CD44+/CD24−/low CSC population was collected for further experiments.
3
Isolation and Characterization of Breast Cancer Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MDA-MB231 cells were starved for 24 h in DMEM/F12 without FBS and dissociated into single-cell suspensions using 2.5% trypsin followed by centrifugation at 300 g for 5 min. Cell pellets were resuspended in 40 μl of PBE suspension buffer (for approximately 1× 107 cells) and incubated with CD24 microbeads (CD24 Microbead Kit; Miltenyi Biotec, Bergisch Gladbach, Germany) followed by a magnetic separation. CD24− cells were collected, washed, and incubated with CD44 microbeads (Miltenyi Biotec) at 4°C for 15 min. After washing and resuspension in 500 μl of PBE buffer, magnetic separation was used for enrichment of CD44+/CD24− cells. To detect the effect of IL-6 on STAT3 signaling in BCSCs and non-BCSCs, before isolation MDA-MB-231 cells were cultured in DMEM for 24 h and then cultured again in DMEM/F12 containing 20 ng/ml of IL-6.
4
CD24+ Spinal Cord Cell Isolation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Neural Dissociation System 6 (CHI Scientific, Maynard, MA, USA) was used to enzymatically disassemble the spinal cord samples. The resulting cells were washed twice with 0.2 µm filtered solution containing phosphate-buffered saline (PBS), pH 7.2, 0.5% bovine serum albumin (BSA), and 2 mM EDTA. The cells were positively selected using four LS separation columns with a CD24 MicroBead Kit on QuadroMACS™ Separator (all from Miltenyi Biotec, Bergisch Gladbach, Germany).
5
Isolation of Breast Cancer Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mammospheres were harvested and enzymatically dissociated into single-cell suspension. Cell suspension was centrifuged at 300 × g for 10 minutes, and the cell pellet was resuspended in 40 μl suspension buffer (~107 total cells). The cells were then incubated with CD24 Microbead Kit and CD44 Microbeads (Miltenyi Biotec, Bergisch Gladbach, Germany) for 15 minutes in refrigerator (4°C), then washed and resuspended in 500 μl buffer, followed by magnetic separation. The CD44+/CD24− cells were then collected as the BCSCs.
6
CD24 and CD44 Expression in MCF-7 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Here, 1 × 107 MCF-7 cells were grown with primary antibody against CD24 following the manufacturer’s protocol for the CD24 Microbead Kit (Miltenyi Biotec, Bergisch Gladbach, Germany). Labeled cells were then incubated with goat anti-mouse IgG microBeads (Miltenyi Biotec) and magnetically separated with MiniMACS columns (Miltenyi Biotec). Acquired CD24− cells were incubated with CD44 microbeads (Miltenyi Biotec) at 4 °C for 15 mins. Cells were again washed and magnetically separated.
7
Isolation of CSC-like Subpopulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Isolation of the CSC-like subpopulation was conducted via magnetic-activated cell sorting (MACS) separation, which involved a MACS Column and MACS separator (Miltenyi Biotec, Bergisch Gladbach, Germany). The parental MDA-MB-231 cells (pMDA) were filtered using a 70-µm cell strainer (BD, Franklin Lakes, NJ, USA) to obtain single cell suspension prior to cell sorting. MACS separation was conducted according to the protocol provided by manufacturer. Briefly, the 24- subpopulation was first isolated via CD24 MicroBead kit (Miltenyi Biotec, Bergisch Gladbach, Germany) by positive depletion of CD24-expressing cells, followed by positive selection of CD44-expressing cells using anti-CD44-PE (Biolegend, San Diego, CA, USA) and anti-PE MicroBead (Miltenyi Biotec, Bergisch Gladbach, Germany). This resulted in the acquisition of the sorted CD24−/CD44+ CSC-like subpopulation (sMDA).
8
Isolation and Characterization of CD24-Defined Cell Subpopulations
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The different cell subpopulations (CD24+ and CD24−) were sorted from parental MDA-MB-231 cells using CD24 MicroBead Kit (MACS Miltenyi Biotec) following manufacturer’s instructions. Briefly, the CD24+ cells were indirectly magnetically labeled with CD24-Biotin antibodies and Anti-Biotin MicroBeads. Then the cell suspension was loaded onto a MACS Column, which is placed in the magnetic field of a MACS Separator. The magnetically labeled CD24+ cells were retained within the column. The unlabeled cells (CD24− cells) ran through; this cell fraction was thus depleted of CD24+ cells. After removing the column from the magnetic field, the magnetically retained CD24+ cells can be eluted as the positively selected cell fraction. The purity of the sorted populations was verified by Flow Cytometry.
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!