Human Thrombopoietin R/Tpo R PE-conjugated Antibody

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Heparinized bone marrow (BM) and peripheral blood (PB) cells (106 leukocytes per tube) were incubated with various combinations of mAb (Supplementary Table S3) for 15 minutes. Then, erythrocytes were lysed in FACS-Lysing-Solution (BD Biosciences, San José, CA, USA). Washed cells were acquired on a FACSCalibur (BD Biosciences) and analyzed by FlowJo software (TreeStar, Ashland, OR, USA)

Protocol tips
Heparinized bone marrow (BM) and peripheral blood (PB) cells (106 leukocytes per tube) were incubated with various combinations of mAb (Supplementary Table S3) for 15 minutes. Then, erythrocytes were lysed in FACS-Lysing-Solution (BD Biosciences, San José, CA, USA). Washed cells were acquired on a FACSCalibur (BD Biosciences) and analyzed by FlowJo software (TreeStar, Ashland, OR, USA)

Publication protocol

Heparinized bone marrow (BM) and peripheral blood (PB) cells (106 leukocytes per tube) were incubated with various combinations of mAb (Supplementary Table S3) for 15 minutes. Then, erythrocytes were lysed in FACS-Lysing-Solution (BD Biosciences, San José, CA, USA). Washed cells were acquired on a FACSCalibur (BD Biosciences) and analyzed by FlowJo software (TreeStar, Ashland, OR, USA) as reported (Blatt et al., 2014; Herrmann et al., 2012, 2014). Antibody-reactivity was controlled using isotype-matched control-antibodies. The gating-strategy to detect CD34+/CD38− stem cells and CD34+/CD38+ progenitor cells is shown in Supplementary Figure S1. Flow cytometric results were expressed as ratio of median fluorescence intensities (MFI) obtained with specific mAb and control mAb (MFI mAb: MFI control mAb). The MFI ratio was graded according to a published score (Blatt et al., 2014; Herrmann et al., 2012, 2014): MFI ratio >100: +++, MFI ratio 10.01-100: ++, MFI ratio 3.01-10: +, MFI ratio 1.51-3: +/−, MFI ratio ≤1.50: –. In a separate set of experiments, cell lines were incubated with imatinib, nilotinib, ponatinib, BEZ235, RAD001, RDEA119, PD0325901 (each 0.1-1 µM), or pimozide (1-10 µM) at 37°C for 24 hours before stained with a PE-conjugated CD25 mAb or IL-1RAP mAb.

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Flow cytometry Anti-bodies Human - CD110/Thrombopoietin R using Human Thrombopoietin R/Tpo R PE-conjugated Antibody from R&D Systems.

Manufacturer protocol

Download the product protocol from R&D Systems for Human Thrombopoietin R/Tpo R PE-conjugated Antibody below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Flow cytometry Anti-bodies Human - CD110/Thrombopoietin R using Human Thrombopoietin R/Tpo R PE-conjugated Antibody from R&D Systems. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.