Cd4 pc5

Manufactured by BD

Sourced in United States

The CD4 PC5 is a lab equipment product designed for cell analysis and flow cytometry applications. It is used to detect and quantify CD4-positive cells in a sample. The core function of the CD4 PC5 is to provide accurate and reliable measurement of CD4 expression on cells.

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Lab products found in correlation

Live dead aqua, by Thermo Fisher Scientific (3 mentions) Cd3 apc h7, by BD (3 mentions) Cd8 v450, by BD (3 mentions) Pd 1 clonem1h4 a488, by BD (2 mentions) Lsrfortessa, by BD (1 mentions) Fc500, by Beckman Coulter (1 mentions) Igg1 pe, by BD (1 mentions) Igg1 fitc, by BD (1 mentions) Pd l1 fitc, by BD (1 mentions) Pd 1 pe, by BD (1 mentions) Ctla 4 pe, by BD (1 mentions)

4 protocols using cd4 pc5

PD-1 Expression on T Cell Subsets

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PBMCs obtained at A5321 study entry were stained with the following monoclonal antibodies to evaluate surface PD-1 expression: CD3 APC-H7 (catalog 560176), CD4 PC5 (catalog 555348), CD8 V450 (catalog 560347), PD-1 (clone M1H4), A488 (catalog 557860) (all from BD Biosciences), and LIVE/DEAD Aqua (Invitrogen, Thermo Fisher Scientific). Cells were fixed in 1% paraformaldehyde and analyzed using a BD LSRFortessa (BD FACSDiva software) within 24 hours after staining. Lymphocytes were identified based on size and granularity. The lymphocyte population was filtered through side scatter area versus side scatter height histogram to eliminate doublets from the analysis. Single cells were analyzed using LIVE/DEAD Aqua dye exclusion, and then CD4⁺ and CD8⁺ populations were defined based on dual expression with CD3. These 2 populations were plotted against PD-1. Fluorescence minus one controls were used to define the PD-1⁺ T cell populations.

Stevenson E.M., Ward A.R., Truong R., Thomas A.S., Huang S.H., Dilling T.R., Terry S., Bui J.K., Mota T.M., Danesh A., Lee G.Q., Gramatica A., Khadka P., Alberto W.D., Gandhi R.T., McMahon D.K., Lalama C.M., Bosch R.J., Macatangay B., Cyktor J.C., Eron J.J., Mellors J.W, & Jones R.B. (2021). HIV-specific T cell responses reflect substantive in vivo interactions with antigen despite long-term therapy. JCI Insight, 6(3), e142640.

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Characterizing PD-1-Expressing T Cells in HIV

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Peripheral blood mononuclear cells (PBMC) obtained from at least four
time points (pre-ART, 1, 4, and, where available, 6–15, years on-ART)
were stained with the following mAbs: CD3 APC-H7, CD4 PC5, CD8 V450, PD-1 (clone
M1H4) A488 (all from BD Biosciences, San Diego, CA), and Live/Dead Aqua
(Invitrogen, Grand Island, NY). Cells were fixed in 1% paraformaldehyde, and
analyzed using a BD LSR Fortessa (FACSDiva) within 24 hours after staining.
Fluorescence minus one (FMO) controls were used in setting gates for positive
events. Cluster and population analysis of flow cytometry plots with a mean
fluorescence intensity (MFI) of at least 5-fold the FMO control MFI determined
the PD-1^HI T cell subset (Suppl Figure 1). Non-specific
cellular staining by the PD-1 antibody was confirmed by analyzing HEK-293 cell
lines ± transfection with plasmid pBudCE4.1/iRFP-hPD-1 as
published^{[31 (link)]} (Suppl Figure 2).

MACATANGAY B.J., GANDHI R.T., JONES R.B., MCMAHON D.K., LALAMA C.M., BOSCH R.J., CYKTOR J.C., THOMAS A.S., BOROWSKI L., RIDDLER S.A., HOGG E., STEVENSON E., ERON J.J., MELLORS J.W, & RINALDO C.R. (2020). T cells with high PD-1 expression are associated with lower HIV-specific immune responses despite long-term antiretroviral therapy. AIDS (London, England), 34(1), 15-24.

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Phenotyping Cytotoxic T Cells via Flow Cytometry

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The antibodies used for CTL phenotyping (anti-CD3-ECD, CD4-PC5, CD8-PC7, PD-1-PE, CTLA-4PE, PD-L1 FITC; IgG1-FITC, IgG1-PE) were all obtained from BD Biosciences. To investigate competitive binding of staining antibody with blocking PD-1 mAb, flow cytometric analysis was performed using two different clones of staining antibody after 48 hrs: primary PD-1 (MIH4), secondary IgG1 FITC and PD-1 conjugated with PE.
Positive staining for PD-1 and PD-L1 was determined by comparing with the respective isotype control. The cell suspensions were incubated with mAb for 30 min at 4°C and then washed twice in PBS. PD-1 mAb-treated co-cultures were incubated with an anti-CD107a antibody and degranulation assay was performed.29 (link) The cells were acquired on an FC 500 (Beckman Coulter) and analyzed by the CXP analysis software.

Kumar R., Yu F., Zhen Y.H., Li B., Wang J., Yang Y., Ge H.X., Hu P.S, & Xiu J. (2017). PD-1 blockade restores impaired function of ex vivo expanded CD8+ T cells and enhances apoptosis in mismatch repair deficient EpCAM+PD-L1+ cancer cells. OncoTargets and therapy, 10, 3453-3465.

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PD-1 Expression on CD4+ and CD8+ T Cells

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PBMCs obtained at A5321 study entry were stained with the following monoclonal antibodies to evaluate surface PD-1 expression: CD3 APC-H7, CD4 PC5, CD8 V450, PD-1 (clone M1H4) A488 (all from BD Biosciences, San Diego, California, USA), and Live/Dead Aqua (Invitrogen, Grand Island, New York, USA). Cells were fixed in 1% paraformaldehyde, and analyzed using a BD LSR Fortessa (FACSDiva) within 24 hours after staining. Lymphocytes were identified based upon size and granularity. The lymphocyte population was filtered through side scatter area vs. side scatter height histogram to eliminate doublets from the analysis. Single cells were analyzed using Live/Dead Aqua dye exclusion and then CD4 + and CD8 + populations were defined based on dual expression with CD3. These two populations were plotted against PD-1. Fluorescence minus one (FMO) controls were used to define the PD-1 + T-cell populations.

Stevenson E.M., Ward A.R., Truong R., Thomas A., Huang S., Dilling T.R., Terry S., Bui J., Mota T.M., Danesh A., Lee G.Q., Gramatica A., Khadka P., Alberto W.D.C., Gandhi R.T., McMahon D., Lalama C.M., Bosch R.J., Macatangay B., Cyktor J.C., Eron J.J., Mellors J.W., & Jones R.B. (2020). HIV-specific T-cell responses reflect substantive in vivo interactions with infected cells despite long-term therapy.

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