BMDCs were stimulated with Rv2005c (10 μg/mL) for 24 h. The stimulated cells were harvested and washed with PBS. The BMDCs were stained with PE-conjugated anti-CD40, anti-CD80, anti-CD86, anti-MHC class I and anti-MHC class II and with FITC-conjugated CD11c (eBioscience) for 30 min at 4 °C. The cells were washed with PBS and suspended in 250 μL PBS. The fluorescence was measured by flow cytometry, and the data were processed using FlowJo software.
Pe conjugated anti cd40
Manufactured by Thermo Fisher Scientific
PE-conjugated anti-CD40 is a fluorescent-labeled antibody that binds to the CD40 cell surface receptor. It is used in flow cytometry applications to identify and quantify CD40-expressing cells.
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Lab products found in correlation
Anti cd80, by Thermo Fisher Scientific (1 mentions)
Anti cd86, by Thermo Fisher Scientific (1 mentions)
Anti mhc class 1, by Thermo Fisher Scientific (1 mentions)
Anti mhc class 2, by Thermo Fisher Scientific (1 mentions)
Fitc conjugated cd11c, by Thermo Fisher Scientific (1 mentions)
Lsrfortessa cell analyzer, by BD (1 mentions)
Anti cd8 pacific blue, by BD (1 mentions)
Pe conjugated anti cd80, by Thermo Fisher Scientific (1 mentions)
Pe conjugated anti cd86, by Thermo Fisher Scientific (1 mentions)
Anti cd4 fluorescein isothiocyanate fitc, by BD (1 mentions)
Brefeldin a, by BD (1 mentions)
2 protocols using pe conjugated anti cd40
1
Characterization of BMDC Activation
2
Endogenous CD4 T cell responses and B cell costimulatory molecules in infected chimera
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Endogenous CD4 T cell responses in infected chimera were analyzed as described previously (Nothelfer et al., 2015) . Briefly, splenocytes were either restimulated with bone marrow-derived dendritic cells (BMDCs) pulsed with fixed parasites or phorbol 12-myristate 13-acetate (PMA)/ionomycin in the presence of Brefeldin A (BD Biosciences). Cells were then stained with anti-CD4-fluorescein isothiocyanate (FITC), anti-CD8 Pacific Blue, anti-IFN-g-allophycocyanin, and anti-IL-10-phycoerythrin (BD Biosciences). 350,000 cells were acquired on a BD LSRFortessa cell analyzer (Becton Dickinson), and analysis was performed using FlowJo software (Tree Star).
The expression of costimulatory molecules by B cells from infected chimeric mice was assessed using the following antibodies: FITC-conjugated anti-major histocompatibility complex class II (MHCII) (BD Biosciences), eFluor 450-conjugated anti-CD19 (eBioscience), PE-conjugated anti-CD40 (eBioscience), PE-conjugated anti-CD80 (eBioscience), and PE-conjugated anti-CD86 (eBioscience). Cells were acquired with a BD LSRFortessa cell analyzer (Becton Dickinson), and analysis was performed using FlowJo software (Tree Star).
The expression of costimulatory molecules by B cells from infected chimeric mice was assessed using the following antibodies: FITC-conjugated anti-major histocompatibility complex class II (MHCII) (BD Biosciences), eFluor 450-conjugated anti-CD19 (eBioscience), PE-conjugated anti-CD40 (eBioscience), PE-conjugated anti-CD80 (eBioscience), and PE-conjugated anti-CD86 (eBioscience). Cells were acquired with a BD LSRFortessa cell analyzer (Becton Dickinson), and analysis was performed using FlowJo software (Tree Star).
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