rev: October 14, 2004
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Human Cytokine/Growth Factor/Chemokines Sample Spec sheets (subject to change without notice):
Description: CD40 ligand (CD40L/TRAP) is a type II membrane protein which effectuates the helper function of T cells on resting B cells. Binding of CD40L to its receptor , CD40, on B cells indices B cell proliferation, adhesion and class switching. This recombinant human soluble CD40 Ligand (sCD40L/TRAp) is a 16.3 kda protein containing 149 amino acid residues comprising the receptor binding TNF-Like domain of CD40L.
Package Size: 50 micrograms $231.00 $219.00/package 2+
Mini-Pack: cat#RDI-3102p $94.00/10ug
-see bulk below
Supplied: Sterile filtered & lyophilized from 10mM Na phosphate pH 7.5.
Source: E. Coli
Purity: >98% by SDS-Page and HPLC analysis Endotoxin level is less than
0.1ng per ug (1EU/ug) of sCD40L/TRAP.
Reconstitution: The lyophilized sCD40L is soluble in most aqueous buffers. The lyophilized protein should be reconstituted in sterile DI water or 5-10mM sodium phophate pH7.5 to a maximum concentration of 1mg/ml. Allow to set at least 30 minutes at 4 DEG C, tighten cap and rotate to reconstitute any material on sides of vial. If desired, you can centrifuge at 1000 rpm for 1 minute to concentrate volume in bottom of vial. This solution can be diluted into other buffered solutions containing carrier protein or medium and stored at 4 DEG C for up to 1 week or at -20 DEG C for future use.
Note:If this material is not to be used for coating or labeling (or other applications requiring carrier free material), the addition of 0.1% BSA to dilution buffers or dilution in sterile culture media is recommended to enhance stability and minimize absorption to vial. Ideally, maintain stock >10ug/ml.
Storage: The lyophilized powder is stable for a few weeks at room temperature but it is best stored desiccated at -20 DEG C. Reconstituted sCD40L should be stored in working aliquots at -20DEG C. AVOID FREQUENT FREEZE THAW CYCLES.
Assay#1: Determined by the stimulation of IL-12 inducation of human PBMC's
Assay#2:The biological activity of this sCD40L/TRAP was determined by the
stimulation of IL-8 prodcution by human peripheral blood mononuclear cells.
The expected ED50 for this effect is 5-10ng/ml. Note:Results will vary wth
Sequence: MQKGDQNPQI AAHVISEASS KTTSVLQWAE KGYYTMSNNL VTLENGKQLT
VKRQGLYYIY AQVTFCSNRE ASSQAPFIAS LWLKSPGRFE RILLRAANTH
SSAKPCGQQS IHLGGVFELQ PGASVFVNVT DPSQVSHGTG FTSFGLLKL
Precautions:For In vitro research Use Only. Not for use in or on humans or animals or for diagnostics. It is the responsibility of the user to comply with all local/state and Federal rules in the use of this product. We are not responsible for any patent infringements that might result with the use of or derivation of this product.
cat# RDI-3102-245X $2500.00 $2250/mg 2 +
Description: Recombinant Human sCD40L is produced in E.Coli as a non-glycosylated, polypeptide chain containing 149 amino acids and having a molecular mass of 16,308 Dalton. rHusCD40L is purified by proprietary chromatographic techniques.
Appearance: Sterile Filtered White lyophilized (freeze-dried) powder from 10mM Sodium Phosphate pH7.5
Endotoxin: Less than 0.1 ng/µg (1EU/µg) of rHusCD40L.
Formulation: Lyophilized from a sterile concentrated solution (1mg/ml) with 10mM Sodium Phosphate pH=7.5.
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.
Solubility: It is recommended to reconstitute the lyophilized rHusCD40L in sterile 18M-cm H2O not less than 100µg/ml, which can then be further diluted to other aqueous solutions.
Stability: Lyophilized rHusCD40L although stable at room temperature for 3 weeks, should be stored desiccated below -18 C. Upon reconstitution rHusCD40L 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.
BiologicalActivity: rHusCD40L is fully biologically active when compared to standard. The ED50, as determined by the dose-dependent stimulation of IL-12 & IL-8 induction by PMB (peripheral Mononuclear) cells was found to be 10-100ng/ml.
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 sCD40L as a Reference Standard.
Precautions: For In vitro research for use in or on humans or animals or for diagnostics. It is the responsibility of the user to complywith all local/state and Federal rules in the use of this product. We are not responsible for any patent infringements that might result with the use of or derivation of this product.
Background: Human CD154 (CD40 Ligand) is a member of the tumor necrosis factor
family and is expressed on the surface of activated T cells. Interaction
of CD154 and CD40 is essential for isotype switching in B cells. Known genetic
defects that alter this interaction lead to impaired immune system function.
CD154 has been shown to be hyperexpressed by B and T cells in SLE patients.
CD154 has been reported to be expressed on vascular endothelial cells, smooth
muscle cells and macrophages indicating a possible role for the CD40-CD154
immunoregulatory signaling mechanism during inflammation and immunity in
cat# RDI-CD154FU-AG Price: $562.00/25ug
package: 25 micrograms (preservative free) in 63ul (0.4mg/ml) 50mM sodium
phosphate pH7.5, 1000mM Potassium Chloride, 150mM NaCl.Product was filtered
through 0.1um and vialed under aseptic conditions. Tighten Cap and centrifuge
1 minute at 1000 rpm to concentrate material in vial prior to use.
Cell Line: murine myeloma cell line P3x63Ag8.653
Mw: a soluble molecule consisting of the extracellular domain (213aa) of human CD154 fused to the extracellular domain (167aa) of murine CD8 alpha
Activity: This human CD154-muCD8 binds to CD40 and blocks binding of anti-CD40 monoclonal antibody. -can be used in histochemistry (frozen sections) and flow cytometry
Storage: 2-5 DEG C (DO NOT FREEZE) aliquot and store in working aliquots.
Protocol: Brifely, 500K cultured Raji cells per tube were washed and incubated 45 minutes in ice with 80ul of CD154-muCD8 at a 5ug/ml concentration. Cells were washed two times and incubated with 50ul of secondary reagent anti-mouse CD8-PE (cat#RDI-MCD8A-53PE $406.00/120T after which they were washed three times, fixed and analyzed by FACS. Cells stained positive with a mean shift of 1.71log10 fluorescent units when compared to a buffer control. Binding was blocked when reagent was preincubated 30 minutes with anti-CD154 (clone 24-31)at a concentartion of 50ug/ml.
For In Vitro Research Use Only.
also available: conjugated to Biotin cat#RDI-CD154FUBT $688.00/25ug
PE cat#RDI-CD154FUPE $938.00/50 tests
see also rec human sCD31 (on special request), sCD54, SCD62E, sCD62P, sCD62L, sCD106(VCAM)
Goat anti-human sCD40-Ligand:Biotin conjugated (antigen aff pure, conjugated to Biotin, lyophilized, carrier free)
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