De-identified UCB was obtained from New York Blood Center. CD34 + HSPCs were isolated by density gradient centrifugation (Lymphoprep, SepMate-50 tubes, Stemcell Tech), followed by magnetic separation on an AutoMACS Pro using microbeads conjugated with anti-CD34 antibodies (Miltenyi). CD34 + cells were cultured overnight in StemSpan SFEM II (Stemcell Tech.) supplemented with 10% fetal bovine serum, penicillin/streptomycin, L-glutamine, 2-mercaptoethanol, 1 μM SR-1 (Stemcell Tech.), 100 ng/mL SCF, 40 ng/mL FLT3L, 50 ng/mL TPO, 20 ng/mL IL3, 20 ng/mL IL6, and 15 ng/mL GM-CSF (Peprotech Inc). Subsequently, cells were sequentially transduced by spinoculation at 1500 RPM at 37 C for 90 min on retronectin-coated plates loaded with pTRIP-MND-GFP (first transduction) or pCL20.MSCV.mir30.PGK.mCherry (second transduction) lentiviral particles in culture media in the presence of 10 μg/mL polybrene. Subsequently, mCherry + GFP + cells were sorted and plated onto semi-solid methylcellulose media (Methocult H4230, Stemcell Tech) supplemented with 5 U/mL EPO, 10 ng/mL IL-3, 5 ng/mL SCF, 5 ng/mL GM-CSF. Colonies were enumerated 12–14 days after plating.
Anti cd34 antibody
Manufactured by Miltenyi Biotec
Sourced in United States, Germany
Anti-CD34 antibodies are a type of laboratory reagent used for the detection and isolation of CD34-positive cells. CD34 is a cell surface marker expressed on hematopoietic stem and progenitor cells. These antibodies can be used in various analytical and cell separation techniques, such as flow cytometry and magnetic cell sorting.
Automatically generated - may contain errors
Lab products found in correlation
Sepmate 50 tube, by STEMCELL (1 mentions)
Interleukin 6 (il 6), by Thermo Fisher Scientific (1 mentions)
Gm csf, by Thermo Fisher Scientific (1 mentions)
Lymphoprep, by STEMCELL (1 mentions)
Methocult h4230, by STEMCELL (1 mentions)
Gm csf, by STEMCELL (1 mentions)
Stemspan sfem 2, by STEMCELL (1 mentions)
Ls column, by Miltenyi Biotec (1 mentions)
Methocult, by STEMCELL (1 mentions)
4 protocols using anti cd34 antibody
1
Lentiviral Transduction of CD34+ HSPCs
2
CD34+ Cell Cytarabine and Phytochemical Study
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CD34 + cells were isolated from MNC by IMCS using anti-CD34 antibodies followed by manufacturer's protocol (Miltenyi Biotec Inc.). These isolated CD34 + cells were treated with various concentrations of Cytarabine (10nM-5000 nM), Hesperidin (0.5-100 μM) and Silibinin (0.5-100 μM) alone as well as in combination with Cytarabine (Hesperidin-25 μM, Silibinin 10 μM) for 24 hours. (Cytarabine-Biobin, Hesperidin-Sigma, Silybinin-Microlabs Ltd.)
3
Separation of Erythroid Progenitors
4
Generation of Mature Mast Cells from CD34+ Progenitors
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human peripheral blood‐derived mast cells were generated from CD34+ progenitor cells according to Schmetzer et al.31 with modifications. Peripheral blood‐derived CD34+ cells were obtained from mononuclear cells by magnetic separation using anti‐CD34 antibody (Miltenyi, Bergisch‐Gladbach, Germany) and LS columns (Miltenyi). The cells were cultured for 6 weeks in a methylcellulose‐based medium (MethoCult, STEMCELL Technologies, Vancouver, BC, Canada) supplemented with 1 µg mL−1 SCF, 0.25 µg mL−1 IL‐6 and 0.025 µg mL−1 IL‐3 to promote differentiation into mast cells. After 6 weeks, the cells were transferred to liquid culture in IMDM media containing 1% Pen Strep, 50 µm 2‐mercaptoethanol, 0.1 µg mL−1 SCF, 0.05 µg mL−1 IL‐6, 1:100 dilution of Insulin‐Transferrin‐Selenium and 0.1% BSA. Cell characterisation and quantification of FcεRI expression confirmed that cells at this point are considered mature mast cells. In order to further upregulate FcεRI expression and, thus, to increase sensitivity to IgE, mast cells were treated with IL‐4 at 10 ng mL−1 for 2 days before use.
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!