Certificate of Analysis and Data Sheet

Source:
E.Coli

Catalog No.
CYT-245

Background:

CD40L or CD154 is a membrane glycoprotein and differentiation antigen expressed on the surface of T-cells. The CD40 ligand stimulates B-cell proliferation and secretion of all immunoglobulin isotypes in the presence of cytokines . CD40 ligand has been shown to induce cytokine production and tumoricidal activity in peripheral blood monocytes. It also costimulates proliferation of activated T-cells and this is accompanied by the production of IFN-gamma , TNF-alpha , and IL2 .

Description :

Recombinant Human CD40L produced in E.Coli is a non-glycosylated, Polypeptide chain containing 149 amino acids and having a molecular mass of 16308 Dalton.
Recombinant CD40L is purified by proprietary chromatographic techniques.

Physical Appearance:

Sterile Filtered White lyophilized (freeze-dried) powder.

Formulation:

CD40L is lyophilized from a sterile concentrated solution (1mg/ml) with 10mM Sodium Phosphate pH=7.5.

Solubility:

It is recommended to reconstitute the lyophilized CD40L in sterile 18M-cm H2O not less than 100ug/ml, which can then be further diluted to other aqueous solutions.

Stability:

Lyophilized CD40L although stable at room temperature for 3 weeks, should be stored desiccated below -18 C. Upon reconstitution CD40L should be stored at 4 C between 2-7 days and for future use below -18 C. For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA).

Please prevent freeze-thaw cycles.

Purity:

Greater than 98.0% as determined by:
(a) Analysis by RP-HPLC.
(b) Anion-exchange FPLC.
(c) Analysis by reducing and non-reducing SDS-PAGE Silver Stained gel.

Amino acid sequence:

The sequence of the first five N-terminal amino acids was determined and was found to be Met-Gln-Lys-Gly-Asp.

Dimers and aggregates:

Less than 1% as determined by silver-stained SDS-PAGE gel analysis.

Biological Activity:

ProSpec's CD40L is fully biologically active when compared to standard. The ED50 as determined by the dose-dependant stimulation of IL-12 & IL-8 induction by PMB (Peripheral Mononuclear) cells was found to be 5-10 ng/ml.

Endotoxin:

Less than 0.1 ng/ug (IEU/ug) of soluble CD40L.

Protein content:

Protein quantitation was carried out by two independent methods:
1. UV spectroscopy at 280 nm using the absorbency value of 1.1 as the extinction coefficient for a 0.1% (1mg/ml) solution. This value is calculated by the PC GENE computer analysis program of protein sequences (IntelliGenetics).
2. Analysis by RP-HPLC, using a standard solution of soluble CD40L as a Reference Standard.

Usage:

Prospec's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.

Gene:

Name:TNFSF5
Synonyms:CD40L, CD40LG, TRAP

Protein synonyms/aliases:

Tumor necrosis factor ligand superfamily member 5 (CD40 ligand)
    (CD40-L)
    (TNF-related activation protein)
    (TRAP)
    (T cell antigen Gp39)
    (CD154 antigen)

Precursor's functional components:

Tumor necrosis factor ligand superfamilymember 5, membrane form
Tumor necrosis factor ligand superfamilymember 5, soluble form .

Protein Family:

Belongs to the tumor necrosis factor family.

Protein Domains:

Domains:
•IPR006052   Tumor Necrosis Factor

Related proteins:

•Recombinant Human BAFF (BLyS)
•Recombinant Human Soluble RANK ligand
•Recombinant Human Tumor Necrosis Factor-alpha
•Recombinant Murine Tumor Necrosis Factor-alpha
•Recombinant Human Tumor Necrosis Factor-beta

Protein links:

Precursor- Protein structure and amino acid sequence:

Latest Publications:

1. Molecular signature of mice T lymphocytes following tolerance induction by allogeneic BMT and CD40-CD40L costimulation blockade.
Transpl Int 2006 Feb; 19(2):146-57
2. Alloimmunity in Primate Heart Recipients with CD154 Blockade: Evidence for Alternative Costimulation Mechanisms.
Transplantation 2006 Jan 27;81(2):255-264
3. Soluble P-selectin and CD40L levels in subjects with prediabetes, diabetes mellitus, and metabolic syndrome-the Chennai Urban Rural Epidemiology Study.
Metabolism 2006 Feb;55(2):237-42
4. Tolerance induction by the blockade of CD40/CD154 interaction in pemphigus vulgaris mouse model.
J Invest Dermatol 2006 Jan;126(1):105-13
5. Inhibition of pemphigus vulgaris by targeting of the CD40-CD154 co-stimulatory pathway: a step toward antigen-specific therapy?
J Invest Dermatol 2006 Jan;126(1):11-3
6. Chorioamnionitis is associated with increased CD40L expression on cord blood platelets.
Thromb Haemost 2005 Dec;94(6):1219-23

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中華民國95年06月06日更新