The largest database of trusted experimental protocols

2 protocols using anti plzf apc

1

Multi-parameter Flow Cytometry Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
Anti-Bcl-2 PE, CD3-FITC, CD8-APC, CD127-BV510, IFNγ-APC, RORγt-BV421, TNF-PE were from BD Biosciences. Anti-CD4-PerCP, CD8-BV510, CD127-PECy7, CD161-BV421, -FITC, and -PE, IFNγ-APC, Ki-67-AF488 and -BV510, TNF-BV421, and Vα7.2 PECy7 were from Biolegend. Anti-PLZF APC was from R&D Systems. CellTrace Violet (CTV) cell proliferation kit, Live/Dead Aqua and Near Infrared fixable cell stain were from Life Technologies. MR1-5-OP-RU tetramer -APC, -BV421, and -PE were from NIH Tetramer Core Facility, Emory University. Staining with the MR1 5-OP-RU tetramers was performed for 40 min at room temperature (RT) (6 (link)) before proceeding to the surface and intracellular staining with other mAbs. Cell surface and intracellular staining for TFs, cytokines, and cytotoxic molecules were performed as previously described (10 (link)). Samples were acquired on an FACS Canto II (BD Biosciences) equipped with 405, 488, and 633 nm lasers or CytoFLEX (Beckman Coulter) equipped with 405, 488, and 638 nm lasers. Data including the compensation platform were analysed using FlowJo v.10 (BD Biosciences).
+ Open protocol
+ Expand
2

Detailed Phenotyping of Immune Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Anti-CD14 and anti-CD19 APC-H7 (both at 1:100), anti-CD25 PECy7 (1:50), anti-CD62L V450 (1:50), anti-CD127 PECy7 (1:100), anti-CD161 FITC (1:10), anti-IFNγ V450 (1:200), anti-Ki67 FITC (1:10) were from BD Biosciences. Anti-CD3 ECD (1:100) and anti-CD8β PE (1:50) were from Beckman Coulter. Anti-Vα7.2 FITC (1:10) and PE (1:20), anti-CD45 Alexa Fluor 700 (1:400), anti-CD45RO PECy7 (1:50), anti-CCR9 Alexa Fluor 647 (1:20), anti-Ki67 Brilliant Violet 421 (1:20), anti-CCR7 Brilliant Violet 421 (1:20) were from BioLegend. Anti-CD161 PerCP-Cy5.5 (1:20), anti-IL-22 PerCP-eFluor 710 (1:50) were from eBioscience. Anti-IL-18R PE (1:20), and anti-PLZF APC (1:10) were from R&D systems. Anti-CD4 Qdot 655 (1:400), anti-CD8α Qdot 605 (1:800), anti-CD45 Qdot 705 (1:100), live/dead aqua (1:100) and near infrared fixable (1:400) cell stain were from Invitrogen. Cell surface staining was performed using directly conjugated antibodies and fixed in Cytofix/Cytoperm (BD Biosciences). Intracellular staining was performed using the appropriate mAb’s in Perm/Wash (BD Biosciences). Samples were acquired on an LSRII flow cytometer (BD Biosciences) equipped with 405, 488, 532 and 647 nm lasers. Single-stained polystyrene beads (BD Biosciences) were used for compensation purposes. Software-based compensation was performed using the compensation platform in FlowJo software version 9.5 (Tree Star).
+ Open protocol
+ Expand

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

Sign up now

Revolutionizing how scientists
search and build protocols!