Dynabeads™ M-280 streptavidin Sheep anti-rabbit and CD31 antibody were mixed in 1:10 ratio and incubated at 4 °C for overnight. ECs differentiated from pEpiSCs were washed with Dulbecco′s phosphate-buffered saline (D-PBS). The cells were detached in 2 mM of ethylene-diamine-tetraacetic acid (EDTA) for 15 min at 39 °C and then centrifuged at 300× g for 3 min. Collected cells were suspended in 1% bovine serum albumin-DMEM medium. Magnetic beads labeled with CD-31 antibody were washed three times with 1% BSA/DMEM using PolyATtract® system 1000 magnetic separation stand (Promega, Z5410). Suspended cells in 1% BSA/DMEM and magnetic beads labeled with the antibody were mixed and then rotated at room temperature for 1 h. Then, the mixtures were washed with 1% BSA/DMEM to remove unlabeled cells using a magnetic stand five times. Sorted ECs were seeded on culture plates coated with 0.2% gelatin for the primary culture. EGM-2, M199 supplemented with 20% of FBS, 30 μg/mL of endothelial cell growth supplement (ECGS) and 100 μg/mL of heparin were used for the culture medium of sorted ECs.
Polyattract system 1000 magnetic separation stand
Manufactured by Promega
Sourced in United States
The PolyATtract® system 1000 magnetic separation stand is a laboratory equipment designed for the isolation and purification of polyadenylated (poly(A)) RNA from total RNA samples. It utilizes magnetic beads coated with oligo(dT) to selectively capture the poly(A) RNA, allowing for efficient separation and recovery of the target RNA molecules.
Automatically generated - may contain errors
2 protocols using polyattract system 1000 magnetic separation stand
1
Isolation and Culture of Endothelial Cells
2
Extraction and Quantification of CTXs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A Bead Beater (BioSpec, Bartlesville, USA) was used for the extraction of CTXs. An Allegra X-15R (Beckman Coulter, Brea, USA) centrifuge was used to obtain the microalgal pellets and in the CTXs extraction after using the sonicator. An Eppendorf 5415D (Hamburg, Germany) centrifuge was used in the CTXs extraction after using the bead beater. Magnetic separation was performed using a MagneSphere Technology Magnetic Separation Stand (for 12 0.5-mL or 1.5-mL tubes) and a PolyATtract System 1000 Magnetic Separation Stand (for one 15-mL tube) from Promega Corporation (Madison, USA). Colorimetric measurements were performed with a Microplate Reader KC4 from BIO-TEK Instruments, Inc. (Vermont, USA). Gen5 software was used to collect and evaluate data. Arrays of eight screen printed carbon electrodes (DRP-8x110), a boxed connector (DRP-CAST8X) and a magnetic support (DRP-MAGNET8X) were purchased from Dropsens S.L. (Oviedo, Spain). The arrays consist of 8 carbon working electrodes of 2.5 mm in diameter, each with its own carbon counter electrode and silver reference electrode. Amperometric measurements were performed with a PalmSens potentiostat connected to an 8-channel multiplexer (MUX8) (Houte, The Netherlands). Data were collected and evaluated with the PalmSens PC software.
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!