For RBD- and S2-specific B cell analysis: Surface staining was performed for 20 min in the presence of 1 mg/mL beriglobin (CSL Behring) with the following fluorochrome-conjugated antibodies titrated to their optimal concentrations: CD20-Viogreen (LT20, Miltenyi), CD14 BV570 (M5E2, Biolegend), CD38 BV605 (HB7, Biolegend), CD27 PE (O323, Biolegend), IgD PerCP-Cy5.5 (IA6-2, Biolegend), CD19 PE-Cy7 (SJ25C1, Biolegend) and CD3 APC-Cy7 (UCHT1, Biolegend) and the labelled proteins (S2 PacBlue (0.25 µg), S2 AlexaFluor488 (0.25 µg), RBD Biotin/Streptavidin PE-Vio615 (0.15 µg) and RBD AlexaFluor647 (0.15 µg)).20 22 (link) Zombie Yellow fixable viability staining (Biolegend) was added during the last 5 min of incubation. After staining, the cells were washed once in PBS/BSA, centrifuged and resuspend in PBS/BSA/2 mM EDTA. For analyses of S-I- and S-II-specific T cells and peripheral blood B and T cell subsets, antibody staining was performed as described before20 22 (link) and in online supplemental methods . All samples were measured on a MACSQuant Analyzer 16 (Miltenyi). Instrument performance was monitored using Rainbow Calibration Particles (BD).
Cd14 bv570 m5e2
Manufactured by BioLegend
Sourced in United States
CD14 BV570 (M5E2) is a fluorochrome-conjugated antibody that binds to the CD14 antigen, a glycosylphosphatidylinositol (GPI)-anchored cell surface receptor expressed primarily on monocytes and macrophages. This product can be used for the identification and analysis of CD14-positive cells in flow cytometry applications.
Automatically generated - may contain errors
Lab products found in correlation
Macsquant analyzer 16, by Miltenyi Biotec (1 mentions)
Zombie yellow fixable viability staining, by BioLegend (1 mentions)
Igd percp cy5.5 ia6 2, by BioLegend (1 mentions)
Cd3 apc cy7 ucht1, by BioLegend (1 mentions)
Rainbow calibration particles, by BD (1 mentions)
Beriglobin, by CSL Behring (1 mentions)
Cd38 bv605 hb7, by BioLegend (1 mentions)
Lsrii flow cytometer, by BD (1 mentions)
Ifn γ pe cy7, by BD (1 mentions)
Cd4 pecy5 rpa t4, by BD (1 mentions)
Il 17a bv421, by BioLegend (1 mentions)
Cd3 bv650 okt3, by BioLegend (1 mentions)
2 protocols using cd14 bv570 m5e2
1
Multiparametric Flow Cytometry for SARS-CoV-2 Antibody Analysis
2
Multicolor Flow Cytometry Phenotyping
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After co-culture with stimulator cells, PBMC were harvested, washed in 1× PBS, stained extracellularly, permeabilized, and stained intracellularly, using a 14-color panel containing Yellow Live/Dead viability kit (Invitrogen, Carlsbad, CA, USA) and mAb to CD14-BV570 (M5E2, Biolegend), CD19-BV570 (HIB19, Biolegend), CD3-BV650 (OKT3, Biolegend), CD4-PECy5 (RPA-T4, BD), CD8-PerCP-Cy5.5 (SK1, BD), CD45RA-biotin (HI100, BD), CD62L-APC-A780 (DREG-56, Ebioscience), integrin α4β7-A647 (ACT1; conjugated in house) streptavidin(SAV)-Qdot800 (Invitrogen) CD69-ECD (TP1.55.3, Beckman Coulter), IFN-γ-PE-Cy7 (B27, BD), TNF-α-A700 (MAb11, BD), IL-2-BV605 (MQ1-17H12, Biolegend), IL-17A-BV421 (BL168, Biolegend), and MIP-1β-PE (IC271P, R&D) as previously described (11 (link)). Samples were acquired using a customized LSRII flow cytometer (BD Biosciences) and analyzed using Winlist v7.0 (Verity Software House, Topsham, ME, USA). Absolute numbers of CD3, CD8, CD4 positive cells and CD8 memory subsets cells were calculated by using percentages obtained from flow cytometry analysis related to the absolute number of lymphocytes determined by routine blood count. Responses against S. Typhi were expressed as net percentage of positive cells (i.e., total percentage of positive cells in the presence of S. Typhi-infected targets minus percentage of positive cells in co-cultures with uninfected cells).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!