Cell Signaling Technology

Product Pathways - Growth Factors/Cytokines

Human CD40 Ligand (hCD40L) #3583

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Source

Recombinant human CD40 Ligand (hCD40L) Met113-Leu261 (Accession #P29965) was produced in E. coli at Cell Signaling Technology.

Molecular Characterization

Recombinant hCD40L has a Met on the amino terminus and has a calculated MW of 16,184. DTT-reduced and non-reduced protein migrate as 16 kDa polypeptides. The expected amino terminus MQKGD of recombinant hCD40L was verified by amino acid sequencing.

Purity

>98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant hCD40L. All lots are greater than 98% pure.

Bioactivity

The bioactivity of recombinant hCD40L was determined in a cell proliferation assay using human B cells. The ED50 of each lot is between 0.5-2 μg/ml.

Coomassie Gel

Coomassie Gel

The purity of recombinant hCD40L was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant hCD40L and staining overnight with Coomassie Blue.

Bioactivity

Bioactivity

The proliferation of human B cells treated with increasing concentrations of hCD40L in the presence of 20 ng/ml human IL-4 (#8919) was assessed. After 96 hour treatment with hCD40L, cells were incubated with a tetrazolium salt and the OD450-OD650 was determined.

Western Blotting

Western Blotting

Western blot analysis of extracts from THP-1 cells, untreated or treated with hCD40L for 15 minutes, using Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb #4668 (upper) and SAPK/JNK (56G8) Rabbit mAb #9258 (lower).


Endotoxin

Less than 0.01 ng endotoxin/1 μg hCD40L.

Formulation

With carrier: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2 containing 20 μg BSA per 1 μg hCD40L. Carrier free: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2.

Background

CD40 Ligand (CD40L), a member of the TNF superfamily of ligands, is expressed as either a membrane-bound or soluble homotrimer (1). Both membrane-bound and soluble forms of CD40L have biological activity (1,2). CD40L is expressed primarily on activated T cells; however, mast cells, basophils, and NK cells may also express CD40L (1). CD40L functions as an important T cell co-stimulatory molecule and enhances T-dependent B cell responses (1,2). CD40L binds CD40, which is a receptor expressed on B cells, dendritic cells, and macrophages (1). Binding of CD40L to CD40 leads to the recruitment of TRAF proteins (TRAFs 2,3,5, and 6) resulting in the activation of the JNK and NF-κB pathways (1,2).

  1. van Kooten, C. and Banchereau, J. (2000) J Leukoc Biol 67, 2-17.
  2. Peters, A.L. et al. (2009) Semin Immunol 21, 293-300.

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For Research Use Only. Not For Use In Diagnostic Procedures.

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