rev:    June 8, 2005

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Human Cytokine/Growth Factor/Chemokines Sample Spec sheets (subject to change without notice):


DATA SHEET:  recombinant soluble HUMAN RANK Ligand (sRANKL)

-Description: RANKL and RANK are members of the TNF superfamily of ligands and receptors that play an important role in the regulation of specific immunity and bone turnover. RANK (receptor) was originally identified as a dendritic-cell-membrane protein, which by interacting with RANKL augments the ability of dendritic cells to stimulate naïve T-cell proliferation in a mixed lymphocyte reaction, to promote the survival of RANK + T cells, and to regulate T-cell-dependent immune response. RANKL, which is expressed in a variety of cells including osteoblasts, fibroblasts, activated T-cells and bone marrow stromal cells, is also capable of interacting with a decoy receptor called OPG. Binding of soluble OPG to sRANKL inhibits osteoclastogenesis by interrupting the signaling between stromal cells and osteoclastic progenitor cells, thereby leading to excess accumulation of bone and cartilage. This recombinant human sRANKL is a 20.0 kDa polypeptide comprising the TNF homologous region of RANKL (176 amino acid residues).

Catalog#: RDI-3001

Package Size: 10 micrograms $231.00   $219.00/package 2+

                             -see bulk below

                     

Supplied: Sterile filtered & lyophilized from 0.5x Tris buffer saline pH7.6 OR  from 5mM sodium phosphate   pH7.5 + 75 mM NaCL. No carrier.

Source: E. Coli

Purity: >98% by SDS-Page and HPLC analysis Endotoxin level is less than 0.1ng per ug (1EU/ug) of RANKL.

Reconstitution: The lyophilized RANKL is soluble in most aqueous buffers. The lyophilized protein should be reconstituted to a 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 -20 DEG C for future use. For most in vitro applications, RANKL exerts its biological activity in the concentration range of 0.2 to 20.0 ng/ml.

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 >50ug/ml.

Storage: The lyophilized powder is stable at room temperature but it is best stored desiccated below 0 Deg C. Reconstituted RaNKL should be stored in working aliquots at -20DEG C. AVOID FREQUENT FREEZE THAW CYCLES.

Reactivity: The ED(50) as determined by the dose dependent stimulation of IL-8 production by Human PBMC is less than 10ng/ml.

AA Sequence:

MEKAMVDGSW LDLAKRSKLE AQPFAHLTIN ATDIPSGSHK

VSLSSWYHDR GWAKISNMTF SNGKLIVNQD GFYYLYANIC

FRHHETSGDL ATEYLQLMVY VTKTSIKIPS SHTLMKGGST

KYWSGNSEFH FYSINVGGFF KLRSGEEISI EVSNPSLLDP

DQDATYFGAF KVRDID

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.


Mouse anti-human sRANK-L 500ug lyophilized (coating ab)

cat#RDI-RANKLabmXP  $375.00

Goat anti-human RANK-L (antigen aff pure, lyophilized, no carrier/no preservative)

cat#RDI-RANKLabGP   $250.00/50ug

cat#RDI-RANKLabGP1   $375.00/100ug

Goat anti-human RANK-L:Biotin conjugated (antigen aff pure, conjugated to Biotin, lyophilized, carrier free)-detecting ab

cat#RDI-RANKLabGPB    $250.00/25ug

cat#RDI-RANKLabGPB1  $375.00/50ug


Recombinant Human Soluble Rank Ligand (rHusRANKL) BULK

cat# RDI-3001-334X     $8125.00     $7500/mg 5 +

       RDI-3001-334S      $250.00/10ug

Description: Recombinant Human sRankL is produced in E.Coli as a non-glycosylated, polypeptide chain containing 176 amino acids and having a molecular mass of 20,006 Dalton. rHusRANKL is purified by proprietary chromatographic techniques.

Source: E.Coli

Appearance: Sterile Filtered White lyophilized (freeze-dried) powder from 10mM Tris pH7.6 and 50mM NaCl

Endotoxin: Less than 0.1 ng/ g (1EU/ g) of rHusRANKL.

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-GLU-LYS-ALA-Met.

Dimers and aggregates:   Less than 1% as determined by silver-stained SDS-PAGE gel analysis.

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

Stability: Lyophilized rHusRANKL 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.

Biological Activity: rHusRANKL is fully biologically active when   compared to standard. The activity is determined by its ability to induce osteoclast formation in RAW264.7 cells using a conc. of 5.0-10.0 ng/ml and by dose dependant stimulation of IL-8 production in human PBMC which was found to be <10 ng/ml.

Protein  content: Protein quantitation was carried out by two independent methods:

1. UV spectroscopy at 280 nm

2. Analysis by RP-HPLC, using a standard solution of s Rank Ligand 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 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.


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