1x106 PBMCs were surface stained using the following anti-human antibodies: FITC-conjugated lineage cocktail (BD Bioscience) containing CD3, CD14, CD16, CD19, CD20, CD56, FITC-conjugated anti-CD11c (clone 3.9), FITC-conjugated anti-CD123 (clone 6H6), e450-conjugated anti-CD161 (HP-3G10), APC-e780-conjugated anti-CD127 (eBioRDR5; all eBioscience), VioGreen-conjugated anti-CD45 (clone 5B1), APC-conjugated anti-CD117 (A3C6E2), PE-conjugated anti-CD294 (BM16; all Miltenyi Biotec). At least 4x105 stained PBMCs were acquired on a FACSCantoIITM (BD Bioscience) and analysed using FlowJo7 software (TreeStar). Previous experiments confirmed that cryopreserved PBMCs did not include any basophils and mast cells using this phenotyping approach. Plasma levels of TSLP and IL-33 by ELISA were analysed using commercially available kits (eBioscience and RnD systems respectively). IL-4, IL-5 and IL-13 were measured using established protocols [50 (link), 58 (link)].
Fitc conjugated lineage cocktail
Manufactured by BD
Sourced in United States
The FITC-conjugated lineage cocktail is a laboratory reagent designed for the identification and analysis of specific cell types. It contains a mixture of antibodies that are conjugated to the fluorescent dye FITC (Fluorescein Isothiocyanate), which allows for the detection and quantification of cells expressing lineage-specific markers.
Automatically generated - may contain errors
2 protocols using fitc conjugated lineage cocktail
1
Comprehensive Immune Profiling of PBMCs
2
Identification and Quantification of ILC2s
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We followed the strategy used by Mjösberg et al.15 (link) Whole blood cells were stained with a fluoresceinisothiocyanate (FITC)-conjugated lineage cocktail, phycoerythrin-CY7-conjugated anti-CD127 Ab and phycoerythrin-conjugated anti-CRTH2 Ab, or the respective isotype Abs (all from BD Pharmingen, San Diego, CA, USA), and a cell lineage cocktail comprising antibodies to CD3, CD14, CD16, CD19, CD34, CD123, CD11c, TCRαβ, and TCRγδ expressed on T cells, monocytes, macrophages, B cells, mast cells, dendritic cells, and hematopoietic progenitor cells. We gated on cells lacking lineage markers and examined expression of CD127 and CRTH2 within this population; with all Lineage-CD127+CRTH2+ lymphocytes considered ILC2s (Fig. 1 ). Finally, cell count was performed using a FACSAriaII flow cytometer (BD Biosciences, San Diego, CA, USA). Data were analyzed using FACSDiva software (BD Biosciences). The number of ILC2s is expressed as a percentage of all lymphocytes.
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!