rev:     November 3, 2004

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


DATA SHEET:recombinant Human BDNF

BDNF is a member of the NGF family of neurotrophic growth factors. Like other members of this family, BDNF supports neuron proliferation and survival. BDNF can bind to a low affinity cell surface receptor called LNGFR, which also binds other neurotrophins such as NGF, NT-3 and NT-4. However, BDNF mediates its neurotrophic properties by signaling through a high affinity cell surface receptor called gp145/trkB. BDNF is expressed as the C-terminal portion of a 247 amino acid polypeptide precursor, which also contains a signal sequence of 18 amino acid residue and a propeptide of 110 amino acid residues. Recombinant human BDNF is a 27.0 kDa homodimer of two 119 amino acid subunits linked by strong non-covalent interactions.

Catalog#: RDI-4502

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

                       Mini-Pack:cat#RDI-4502P $95.00/2ug      see bulk below

Supplied: sterile filtered and Lyophilized protein with no additives.

Source: E. Coli

Purity: >96% by SDS-Page analysis. Endotoxin level is less than 0.1ng per ug of BDNF.

Reconstitution: The lyophilized human BDNF is soluble in water and most aqueous buffers. The lyophilized powder should be reconstituted in water to a concentration of 100 ng/ul. Allow to set at least 30 minutes at 4 Deg C, rotate, mix well. If desired, centrifuge for 1-2 minutes at 500- 1000 rpm to concentrate material in vialThis solution can be diluted into water or other buffered solutions or 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 >50ug/ml.

Storage: The lyophilized powder, though stable at room temperature for a few weeks, is best stored dessicated at -20 DEG C. Reconstituted BNDF should be stored in working aliquots at -20 DEG C. Avoid Frequent Freeze- THAW Cycles.

Reactivity:

Assay#1: The ED50 as determined by the dose dependent induction of choline acetyl transferase activity in RAT Basal Forebrain Septal Cultures was found to be 25-50ng/ml.

Assay#2: Determined by its ability to stimulate chick E8 neurite outgrowth. The expected ED50 for this effect is 0.1-1.0ng/ml

AA Sequence:(N terminal met)

M HSDPARRGEL SVCDSISEWV TAADKKTAVD MSGGTVTVLE

     KVPVSKGQLK QYFYETKCNP MGYTKEGCRG IDKRHWNSQC

     RTTQSYVRAL TMDSKKRIGW RFIRIDTSCV CTLTIKRGR



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.


Recombinant Human BDNF (BULK)

cat# RDI-4502-207X      $6000.00/1mg    $5000.00/mg 5+   Larger amounts on quote

       RDI-4502-207X5      $4500.00/0.5mg

Description : Recombinant Human BDNF produced in E.Coli is a homodimer , non-glycosylated, polypeptide chain containing 2 x 119 amino acids and having a total molecular mass of 26,984 Dalton. rHuBDNF is purified by proprietary chromatographic techniques.

Appearance: Sterile Filtered White lyophilized (freeze-dried) powder.

Endotoxin: Less than 0.1 ng/µg (IEU/µg) of rHuBDNF .

Formulation: The protein was lyophilized after dialysis against 20mM sodium citrate buffer pH=5.

Purity: Greater than 96.0% 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-His-Ser-Asp-Pro.

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

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

Stability: Lyophilized rrHuBDNF although stable at room temperature for 3 weeks, should be stored desiccated below -18 C. Upon reconstitution rHuBDNF 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 avoid freeze-thaw cycles.

Biological Activity: This rHuBDNF is fully biologically active when compared to standard. The ED50, calculated by the dose-dependant induction of ACHE (acetylcholine esterase) in rat basal forebrain primary septal culture is 50 ng/ml.

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

1. UV spectroscopy at 280 nm using the absorbency value of 1.6 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 BMP-2 as a Reference Standard.

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.


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