Log In Sign up
The largest database of trusted experimental protocols

Trimeric cd40l

Manufactured by Enzo Life Sciences
Visit Supplier

Trimeric CD40L is a recombinant protein that functions as a costimulatory molecule. It is a trimeric form of the CD40 ligand protein, which plays a key role in the activation of B cells, dendritic cells, and other immune cells.

Automatically generated - may contain errors

Lab products found in correlation

Lsrii flow cytometer, by BD (2 mentions) Phosflow perm buffer 3, by BD (2 mentions) Cytofix fixation buffer, by BD (2 mentions) V450 coupled anti cd19, by BD (2 mentions) Pe cy5 coupled anti cd3, by BioLegend (2 mentions) Fitc coupled anti pp38, by Cell Signaling Technology (2 mentions) Pe coupled anti pp65, by BD (2 mentions) Pe coupled anti perk, by BD (2 mentions) Phytohemagglutinin (pha), by Merck Group (1 mentions) Interleukin 2 (il 2), by Roche (1 mentions) Phytohemagglutinin (pha), by R&D Systems (1 mentions) Interleukin 2 (il 2), by R&D Systems (1 mentions) Ccl19, by R&D Systems (1 mentions) Gm csf, by Miltenyi Biotec (1 mentions)

Spelling variants of this product

CD40L (24 citations)

3 protocols using trimeric cd40l

1

Isolation and Activation of Immune Cells

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
CD4+ T lymphocytes, B lymphocytes, and CD14+ monocytes were positively selected from PBMC using anti-CD4, -CD19, or -CD14 monoclonal antibody (MAb)-coated magnetic beads (Miltenyi Biotech). Immature DC (iDC) were derived from CD14+ monocytes cultured with 1,000 U/ml granulocyte-macroPHAge colony-stimulating factor (GM-CSF; Miltenyi Biotech) and 1,000 U/ml recombinant human interleukin-4 (rhIL-4) for 5 days in AIM-V medium, with additional GM-CSF and rhIL-4 added on day 3. Mature DC (mDC) were derived from iDC by the addition of 0.1 μg/ml trimeric CD40L (Enzo) on day 5 and cultured for an additional 2 days. Prior to coculture, CD4+ T cells and B cells were activated for 48 h with 10 U/ml IL-2 (Roche) and 2 μg/ml phytohemagglutinin (PHA; Sigma) or 1,000 U/ml rhIL-4 and 0.1 μg/ml trimeric CD40L (Enzo), respectively. CD4+ TN or TCM were treated with either 100 nM CCL-19 (R&D Systems) or 10 U/ml IL-2 and 2 μg/ml PHA for 48 h as previously described (12 (link), 15 (link)).
Gerberick A., DeLucia D.C., Piazza P., Alaoui-El-Azher M., Rinaldo C.R., Sluis-Cremer N, & Rappocciolo G. (2021). B Lymphocytes, but Not Dendritic Cells, Efficiently HIV-1 Trans Infect Naive CD4+ T Cells: Implications for the Viral Reservoir. mBio, 12(2), e02998-20.
+ Open protocol
+ Expand
2

Phosphorylation Profiling of Immune Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
For determination of phosphorylated proteins after CD40 stimulation, PBMCs were thawed and rested for 2 h and then either left unstimulated or stimulated with trimeric CD40L (100 ng/ml) (Enzo Life Sciences, Farmingdale, NY) for 15 min, performed concurrently with decoration with V450-coupled anti-CD19 (Catalog#: 560353, BD Biosciences). After incubation with BD Cytofix™ Fixation Buffer (Catalog#: 554655) for 10 minutes at 37°C;, followed by permeabilization in BD Phosflow™ Perm Buffer III (Catalog#: 558050) for 30 minutes on ice, cells were then stained with PE-Cy5-coupled anti-CD3 (BioLegend), FITC-coupled anti-pp38 (Catalog#: 4551, Cell Signaling Technology, Danvers, MA), PE-coupled anti-pp65 (Catalog#: 558423, BD Biosciences), or PE-coupled anti-pErk (Catalog#: 612566, BD Biosciences). The analysis was performed using an LSR II flow cytometer (BD Biosciences) and FlowJo 7.6.2 software (TreeStar, Ashland, OR).
Yu N., Lambert S., Bornstein J., Nair R.P., Enerbäck C, & Elder J.T. (2018). The Act1 D10N Missense Variant Impairs CD40 Signaling in Human B-Cells. Genes and immunity, 20(1), 23-31.
+ Open protocol
+ Expand
3

Phosphorylation Profiling of Immune Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
For determination of phosphorylated proteins after CD40 stimulation, PBMCs were thawed and rested for 2 h and then either left unstimulated or stimulated with trimeric CD40L (100 ng/ml) (Enzo Life Sciences, Farmingdale, NY) for 15 min, performed concurrently with decoration with V450-coupled anti-CD19 (Catalog#: 560353, BD Biosciences). After incubation with BD Cytofix™ Fixation Buffer (Catalog#: 554655) for 10 minutes at 37°C;, followed by permeabilization in BD Phosflow™ Perm Buffer III (Catalog#: 558050) for 30 minutes on ice, cells were then stained with PE-Cy5-coupled anti-CD3 (BioLegend), FITC-coupled anti-pp38 (Catalog#: 4551, Cell Signaling Technology, Danvers, MA), PE-coupled anti-pp65 (Catalog#: 558423, BD Biosciences), or PE-coupled anti-pErk (Catalog#: 612566, BD Biosciences). The analysis was performed using an LSR II flow cytometer (BD Biosciences) and FlowJo 7.6.2 software (TreeStar, Ashland, OR).
Yu N., Lambert S., Bornstein J., Nair R.P., Enerbäck C, & Elder J.T. (2018). The Act1 D10N Missense Variant Impairs CD40 Signaling in Human B-Cells. Genes and immunity, 20(1), 23-31.
+ 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!