Sample collection and study protocols were approved by the Stanford Institutional Review Board (IRB). Samples from treatment naive patients were collected with informed consent and banked by the Stanford Division of Hematology. T-ALL cells were purified from peripheral blood or marrow samples by consulting the patient’s clinical flow phenotype and performing FACS using the following antibodies: anti-CD34:PE clone 582 (BD), anti-CD3:PE-Cy7 clone UCHT1 (BD), anti-CD7:APC clone M-T70 (BD), anti-CD10:APC-Cy7 clone HI10a (Biolegend), anti-CD14:PE-Cy5 clone 61D3 (Invitrogen), anti-CD123:PE-Cy5 clone 6H6 (Biolegend), anti-CD19:PE-Cy5 clone HIB19 (BD), and anti-CD11b:PE-Cy5 clone ICRF44 (BD) (SI Appendix, Fig. S24 ). Purified tumor cells were cultured for 48 h over a monolayer of GFP+ hTERT-immortalized mesenchymal stem cells in the presence of the MYC inhibitor 10058-F4 (Sigma). For analysis of Siglec ligands, the cells were resorted and stained with Siglec-Fc probes as described above using a polyclonal goat anti-human IgG:Brilliant Violet 421 (Jackson ImmunoResearch) secondary antibody.
Anti cd3 pe cy7 clone ucht1
Manufactured by BD
Anti-CD3:PE-Cy7 clone UCHT1 is a fluorescently-labeled monoclonal antibody that binds to the CD3 complex on human T cells. It is suitable for flow cytometry applications.
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Lab products found in correlation
2 protocols using anti cd3 pe cy7 clone ucht1
1
T-ALL Cell Purification and Culture
2
PBMC Isolation and Flow Cytometry
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Peripheral blood mononuclear cells (PBMCs) were isolated from the peripheral blood samples drawn from patients. Briefly, 1×106 cells were washed with phosphate-buffered saline (PBS) and resuspended in fluorescence activated cell sorting (FACS) buffer consisting of PBS and 2% FBS. Cells were then incubated with anti-CD3-PE-cy7 (clone UCHT1; BD Pharmingen), CD19 (20-291) protein-FITC (ACRO Biosystems), or anti-CD19-PE (clone HIB19; BioLegend) antibodies for 30 min. Flow cytometry analysis was performed on BD FACS CantoII (BD, USA).
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