Frozen liver tissue sections from all groups were fixed, blocked and incubated with primary antibody CD8, CD34, CD62L (Abcam, MA) and MECA-79 (Biolegend, CA). Then the tissues were probed with secondary antibodies and counter stained with nuclear stain DAPI. Images were taken using Olympus FV1000 confocal laser-scanning microscope (Olympus America Inc.) and the generated data sets were analyzed using Imaris 8.4 (Bitplane) for 3D co-localization and visual overlap at Emory Integrated Cellular imaging Core.
Cd62l
Manufactured by Abcam
Sourced in United States, United Kingdom
CD62L, also known as L-selectin, is a cell adhesion molecule expressed on the surface of leukocytes. It plays a crucial role in the initial tethering and rolling of leukocytes along the vascular endothelium, facilitating their migration to sites of inflammation or infection.
Automatically generated - may contain errors
Lab products found in correlation
Fv1000 confocal laser scanning microscope, by Olympus (1 mentions)
Meca 79, by BioLegend (1 mentions)
Icam 1, by Abcam (1 mentions)
Vcam 1, by Abcam (1 mentions)
Dna histone complexes, by Merck Group (1 mentions)
Lsrii flow cytometer, by BD (1 mentions)
Cell stimulation cocktail, by Thermo Fisher Scientific (1 mentions)
Brefeldin a, by BD (1 mentions)
Cytofix cytoperm kit, by BD (1 mentions)
Anti ifn γ xmg1, by BioLegend (1 mentions)
Il 17, by Thermo Fisher Scientific (1 mentions)
Cyan adp flow cytometer, by Beckman Coulter (1 mentions)
4 protocols using cd62l
1
Multicolor Immunofluorescence Imaging of Liver
2
Multiplex Biomarker Analysis in Serum
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Various ELISAs were used for detection of serum levels of S100A8/A9, myeloperoxidase (MPO), neutrophil elastase (NE), CXCL8, VCAM-1, ICAM-1, CD62L (Abcam), and DNA-histone complexes (Merck Millipore). For CXCL4, lipocalin, lactoferrin, MMP8, proteinase 3, sCD62P, and sCD40L were determined by LUMINEX xMAP Technology.
3
Phenotyping of Skin-Draining Lymph Node Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Skin-draining lymphnodes (SDLNs) were dissociated and filtered with a 40-mm cellstrainer. Blood cells were depleted of erythrocytes by ammonium chloride lysis and washed before staining. Flow cytometric analysis of immune cell phenotype was carried out by staining with the following fluorochrome-conjugated Abs: CD8 (12-0081-82, eBioscience, USA), CD44 (16-0441-85, eBioscience) and CD62L (ab25282, Abcam) as described previously [31 (link)]. For intracellular IFN-γ assay, 2 × 105 CD8+ T cells were treated with Cell Stimulation Cocktail (eBioscience) for 1 h. Brefeldin A (BD Biosciences) was added for 4 h incubation at 37°C. Cells were then fixed and permeabilized using the Cytofix/Cytoperm kit (BD Biosciences) and stained intracellularly with anti-IFN-γ (XMG1.2, BioLegend) for 30 min at 4°C. The corresponding isotype controls (551954, BD Biosciences) were used for flow cytometry. Data were collected using an LSR II flow cytometer (BD Biosciences). FlowJo 7.6 software is used for the analysis of flow cytometric data.
4
Multiparameter Flow Cytometry of Immune Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Freshly-isolated PBMC, SVF and intrahepatic immune cells were stained with monoclonal antibodies (mAbs) specific for human surface markers (CD3, CD4, CD8, CD45RA, CD69; BD Biosciences, Oxford, UK), CD27 (eBioscience, Hatfield, UK) and CD62L (Abcam, Cambridge, UK). Gating strategy for surface markers is outlined in supplemental Figure 1A.
For intracellular cytokine staining cells were stimulated with 10 ng/ml of phorbal myristate acetate and 1 µg/ml of ionomycin (PMA/I) for 1 hour, followed by the addition of 10 µg/ml of brefeldin A for a further 3 hours. Cells were stained with mAbs specific for human surface markers CD3 and CD8 for 30 minutes. As human CD4 cannot be reliably detected following treatment with PMA, CD4 + T cells were represented by CD3 + CD8 -cells, the vast majority of this population is made up of CD4 + T cells, however it will also contain minor populations of innate lymphocytes. Cells were fixed and permeabilised, then stained with mAbs specific for the cytokines IFN-, TNF-α and IL-10 (BD Biosciences, Oxford, UK) and IL-17 (eBioscience, Hatfield, UK). Gating strategy for intracellular cytokines is outlined in supplemental Figure 1B. Cells were acquired using a CyAn ADP flow cytometer (Beckman Coulter) and analysed with FlowJo software (TreeStar Inc.).
For intracellular cytokine staining cells were stimulated with 10 ng/ml of phorbal myristate acetate and 1 µg/ml of ionomycin (PMA/I) for 1 hour, followed by the addition of 10 µg/ml of brefeldin A for a further 3 hours. Cells were stained with mAbs specific for human surface markers CD3 and CD8 for 30 minutes. As human CD4 cannot be reliably detected following treatment with PMA, CD4 + T cells were represented by CD3 + CD8 -cells, the vast majority of this population is made up of CD4 + T cells, however it will also contain minor populations of innate lymphocytes. Cells were fixed and permeabilised, then stained with mAbs specific for the cytokines IFN-, TNF-α and IL-10 (BD Biosciences, Oxford, UK) and IL-17 (eBioscience, Hatfield, UK). Gating strategy for intracellular cytokines is outlined in supplemental Figure 1B. Cells were acquired using a CyAn ADP flow cytometer (Beckman Coulter) and analysed with FlowJo software (TreeStar Inc.).
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!