We used QBEND-10 monoclonal CD34 antibody (Abcam) with an RQR8 label to detect the effectiveness of 123CL CAR's transduction. T cell immunophenotype was analyzed by the combination of CD3-perCP, CD4-FITC, and CD8-APC monoclonal antibodies. The expression of CD123 and CLL1 in AML blasts was examined with a combination of CD45-mFluor540, CD38-FITC, CD34-APC, CD123-PE, and CLL1-PE/cy7 antibodies to identify diverse cell subpopulations. Positive expression of this antigen is defined as cell expression that exceeds 20%. Coulter Altra flow cytometry (Beckman) was utilized for the detection, while CytExpert or FlowJo7.6.5 was used for the analysis.
Qbend 10 monoclonal cd34 antibody
Manufactured by Abcam
QBEND-10 is a monoclonal antibody that specifically recognizes the CD34 antigen. CD34 is a transmembrane glycoprotein expressed on the surface of hematopoietic stem and progenitor cells. The QBEND-10 antibody is commonly used for the identification and enumeration of CD34-positive cells in various applications.
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3 protocols using qbend 10 monoclonal cd34 antibody
1
Characterizing Effector T Cell Phenotype and Antigen Expression in AML
2
Determining Viral Titer by Limiting Dilution
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The number of transducing units (TU/mL) was determined by the limiting dilution method. Briefly, HEK-293T cells were seeded 12 hr before transduction. Then, 1:10 dilutions of the viral supernatant were prepared and added on top of the cells in complete DMEM + 5 μg/mL Polybrene. Cells were trypsinyzed 72 hr later and labeled with the QBEND-10 monoclonal CD34 antibody (Abcam) before being analyzed by flow cytometry. A dilution corresponding to 2%–20% of positive cells was used to calculate viral titer.
3
Comprehensive Immunophenotyping of CAR-T Cells
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For each analyzed T cell culture, one sample of cells was stained with Alexa-Fluor 647-labeled polyclonal goat anti-mouse IgG (H+L) antibodies (Affinity) to detect CAR-T cells. In addition, we also detected the expression of the CAR with the QBEND-10 monoclonal CD34 antibody (Abcam) labeled with an RQR8 tag. Subsequently, all cells were stained with fluorescein isothiocyanate (FITC)-labeled anti-CD3 antibodies (Abcam).
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