HEK-Blue™ CD40L Cells
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Cat.code:
hkb-cd40
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ABOUT
CD40L responsive NF-κB-SEAP reporter assay
HEK-Blue™ CD40L cells are designed to monitor human CD40L-induced NF-κB stimulation or inhibition. This colorimetric bioassay can be used to screen activatory or inhibitory molecules, such as engineered cytokines and neutralizing antibodies.
HEK-Blue™ CD40L cells respond specifically to recombinant human CD40L and mouse CD40L. Their reliable and consistent performance makes them suitable for release assays of activatory and inhibitory molecules such as Frexalimab, a monoclonal antibody that targets CD40L and prevents its binding to its receptor (see figures).
Key features
- Readily assessable NF-κB-SEAP reporter activity
- Convenient readout using QUANTI-Blue™ Solution
- High sensitivity to human (h) CD40L and mouse (m) CD40L
- Stability guaranteed for 20 passages
Applications
- Therapeutic development
- Drug screening
- Release assay
CD40L is a protein that is primarily expressed on activated T cells and is a member of the tumor necrosis factor superfamily. Of note, CD40L, together with its receptor CD40, plays a pivotal role in cellular and humoral immunity.
Disclaimer: These cells are for internal research use only and are covered by a Limited Use License (See Terms and Conditions). Additional rights may be available.
SPECIFICATIONS
Specifications
CD40L
Human, Mouse
Detection and quantification of CD40L activity
3 ng/ml - 1 µg/ml (hCD40L, mCD40L)
Complete DMEM (see TDS)
Verified using Plasmotest™
Each lot is functionally tested and validated.
CONTENTS
Contents
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Product:HEK-Blue™ CD40L Cells
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Cat code:hkb-cd40
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Quantity:3-7 x 10^6 cells
- 1 ml of Blasticidin (10 mg/ml)
- 1 ml of Zeocin® (100 mg/ml)
- 1 ml Normocin™ (50 mg/ml)
- 1 ml of QB reagent and 1 ml of QB buffer (sufficient to prepare 100 ml of QUANTI-Blue™ Solution, a SEAP detection reagent)
Shipping & Storage
- Shipping method: Dry ice
- Liquid nitrogen vapor
- Upon receipt, store immediately in liquid nitrogen vapor. Do not store cell vials at -80°C.
Storage:
Caution:
Details
Cell line description
HEK-Blue™ CD40L cells were generated by stable transfection of the human embryonic kidney HEK293 cell line with the human CD40 gene and an NF-κB-inducible secreted embryonic alkaline phosphatase (SEAP) construct. The SEAP construct comprises the SEAP reporter gene under the control of the IFN-β minimal promoter, fused to five NF-κB binding sites. Binding of CD40L to its receptor CD40 triggers a signaling cascade leading to the activation of NF-κB and the subsequent production of SEAP. CD40L-CD40 interaction can be monitored by assessing the levels of SEAP using QUANTI-Blue™ Solution, a SEAP detection reagent.
HEK-Blue™ CD40L cells detect human (h) and mouse (m) CD40L. These cells also respond to recombinant hIL-1β and hTNF-α (see figures).
CD40 Ligand background
CD40 Ligand (CD40L), also known as CD154, TRAP, or gp39, is a type II transmembrane glycoprotein belonging to the tumor necrosis factor (TNF) family. It is mainly expressed in CD4+-T cells and interacts with CD40 on antigen-presenting cells to regulate both humoral and cellular immune responses [1-3].
The CD40 cytoplasmic domain binds directly to several TNF receptor-associated factors (TRAFs), and this interaction is thought to initiate CD40 signaling. CD40-mediated signaling results in NF-κB, c-Jun N-terminal kinase (JNK), and p38 mitogen-activated protein kinase (MAPK) activation. CD40L-CD40 interactions are thought to play an important role in the pathogenesis of many diseases [4, 5].
References:
1. Karnell J.L. et al., 2019. Targeting the CD40-CD40L pathway in autoimmune diseases: Humoral immunity and beyond. Adv Drug Deliv Rev. 141:92-103.
2. Laman J.D. et al., 2017. Functions of CD40 and Its Ligand, gp39 (CD40L). Crit Rev Immunol. 37(2-6):371-420.
3. Elgueta R. et al., 2009. Molecular mechanism and function of CD40/CD40L engagement in the immune system. Immunol. Rev. 229, 152–172.
4. Seijkens T. et al., 2013. CD40–CD40L: Linking pancreatic, adipose tissue and vascular inflammation in type 2 diabetes and its complications. Diabetes and Vascular Disease Research. 10: 115 - 122.
5. Daoussis D. et al., 2004. Targeting CD40L: a promising therapeutic approach. Clin Diagn Lab Immunol. 11(4):635-41.
DOCUMENTS
Documents
Technical Data Sheet
Validation Data Sheet
Safety Data Sheet
Certificate of analysis
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