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Cd3 fitc clone hit3a

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CD3 FITC (clone HIT3A) is a fluorescently labeled monoclonal antibody that binds to the CD3 antigen expressed on the surface of T cells. It is used in flow cytometry applications for the identification and enumeration of T cells in biological samples.

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Lab products found in correlation

Cd19 fitc clone hib19, by BD (1 mentions) Cd19 pe clone hib19, by BD (1 mentions) Cd4 pe clone rpa t4, by BD (1 mentions) Acuri c6, by BD (1 mentions) Cd8 apc clone rpa t8, by BD (1 mentions) Cd4 apc clone sk3, by BD (1 mentions) Cd45ra apc h7 clone hi100, by BD (1 mentions) Ccr7 pe cy7 clone 3d12, by BD (1 mentions) Cd27 pe clone m t271, by BD (1 mentions) Facsaria 2 cell sorter, by BD (1 mentions)

2 protocols using cd3 fitc clone hit3a

1

Measuring MET-CAR Transduction Efficacy

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To measure MET-CAR transduction efficacy, cells were collected on day 5 after transduction, washed with PBS containing 1% FBS, and incubated with CD3 and CD19 antibodies for 30 min followed by analysis using a flow cytometer (Acuri C6 + , BD). To determine the subsets of MET-CAR-T cells, CD19+ T cells were gated for  CD4+ and CD8+ populations. To determine programmed cell death protein 1 (PD-1) upregulation in MET-CAR-T cells, non-transduced (NT) or MET-CAR-T cells were co-cultured with MHCC97H cells in 24-well plates (105 cells/well) at a 1:1 ratio for 3 days. At day 0 (before co-culture) and day 3, NT (gated by CD3+) and MET-CAR-T cells (gated by CD19+) were harvested to measure the PD-1 expression using flow cytometry analysis. Antibodies and isotype controls used in the analysis include: CD3 FITC (clone HIT3A, BD), CD19 PE (clone HIB19, BD), CD19 FITC (clone HIB19, BD), CD4 PE (clone RPA-T4, BD), CD8 APC (clone RPA-T8, BD), PD-1 Cy7(Clone EH12.1, BD), IgG2b κ FITC (clone 27-35, BD), IgG1 PE (clone MOPC-21, BD), IgG2b κ PE-Cy7 (clone 27-35, BD), and IgG1 APC (clone P3.6.2.8.1, Invitrogen, San Diego, CA).
Qin A., Qin Y., Lee J., Musket A., Ying M., Krenciute G., Marincola F.M., Yao Z.Q., Musich P.R, & Xie Q. (2023). Tyrosine kinase signaling-independent MET-targeting with CAR-T cells. Journal of Translational Medicine, 21, 682.
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2

Quantifying CD4+ T Cell Subsets

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CD4+ T cell subsets were sorted on a BD FACSAria II cell sorter. The following antibodies were used: CD3-FITC (clone HIT3a, BD #555339), CD4-APC (clone SK3, BD #340443), CD45RA-APCH7 (clone HI100, BD #560674), CCR7-PE-Cy7 (clone 3D12, BD #557648), CD27-PE (clone M-T271, BD #555441) and LIVE/DEAD fixable Aqua (Invitrogen #L34957). Frequencies of memory CD4+ T cell subsets were determined as previously described after the exclusion of dead cells (LIVE/DEAD) (55 (link)). Four CD4+ T cell subsets were defined based on the expression of CD45RA, CCR7 and CD27: naïve (TN: CD45RA+, CCR7+, CD27+), central memory (TCM: CD45RA−, CCR7+, CD27+), transitional memory (TTM: CD45RA−, CCR7−, CD27+) and effector memory (TEM: CD45RA−, CCR7−, CD27−). Integrated HIV DNA was measured in the sorted populations. All subsets for which insufficient number of cells were analyzed (<40,000 sorted cells as measured in the PCR assay) were excluded from the analysis.
Leyre L., Kroon E., Vandergeeten C., Sacdalan C., Colby D.J., Buranapraditkun S., Schuetz A., Chomchey N., de Souza M., Bakeman W., Fromentin R., Pinyakorn S., Akapirat S., Trichavaroj R., Chottanapund S., Manasnayakorn S., Rerknimitr R., Wattanaboonyoungcharoen P., Kim J.H., Tovanabutra S., Schacker T.W., O’Connell R., Valcour V.G., Phanuphak P., Robb M.L., Michael N., Trautmann L., Phanuphak N., Ananworanich J, & Chomont N. (2020). Abundant HIV-infected cells in blood and tissues are rapidly cleared upon ART initiation during acute HIV infection. Science translational medicine, 12(533), eaav3491.
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