Peripheral blood from three ET patients with mutations in CALR underwent Ficoll density gradient separation, immunomagnetic selection for CD34+ cells (Miltenyi Biotech), and fluorescence-activated cell sorting (FACS) (Influx, Becton-Dickinson) using PeCy7-labeled CD34, clone 561 (lot# B257238, BioLegend), APC-labeled CD38, clone HIT2 (lot #B247250, BioLegend) and FITC-labeled CD10, clone HI10a (lot# B254556, BioLegend) antibodies were used to isolate CD34+CD38−, CD34+CD38+, and CD34+CD10+ cell compartments. DNA was extracted from sorted cells (Qiagen) and the VAF of CALR mutations was measured by droplet digital PCR (QX200 Droplet Digital PCR System, Bio-Rad) with primers that specifically detect CALR type 1 mutations (52-bp deletion (p.L367fs*46), CALR type 2 mutations (5-bp TTGTC insertion (p.K385fs*47) or wildtype alleles.
Ficoll density gradient separation
Manufactured by Miltenyi Biotec
Ficoll density gradient separation is a laboratory technique used to isolate and purify specific cell types from a heterogeneous mixture. It utilizes a density gradient medium made of Ficoll, a synthetic polymer, to separate cells based on their density differences. The core function of this method is to enable the efficient and gentle isolation of cells, such as mononuclear cells, from complex biological samples, like blood or tissue.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using ficoll density gradient separation
1
Characterizing CALR Mutant HSCs
2
Characterizing CALR Mutant HSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Peripheral blood from three ET patients with mutations in CALR underwent Ficoll density gradient separation, immunomagnetic selection for CD34+ cells (Miltenyi Biotech), and fluorescence-activated cell sorting (FACS) (Influx, Becton-Dickinson) using PeCy7-labeled CD34, clone 561 (lot# B257238, BioLegend), APC-labeled CD38, clone HIT2 (lot #B247250, BioLegend) and FITC-labeled CD10, clone HI10a (lot# B254556, BioLegend) antibodies were used to isolate CD34+CD38−, CD34+CD38+, and CD34+CD10+ cell compartments. DNA was extracted from sorted cells (Qiagen) and the VAF of CALR mutations was measured by droplet digital PCR (QX200 Droplet Digital PCR System, Bio-Rad) with primers that specifically detect CALR type 1 mutations (52-bp deletion (p.L367fs*46), CALR type 2 mutations (5-bp TTGTC insertion (p.K385fs*47) or wildtype alleles.
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!