rev:   October 19, 2004

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


DATA SHEET:  recombinant Human TUMOR NECROSIS FACTOR-A

-Tumor Necrosis Factor Alpha (TNF-A) is a potent lymphoid factor which exerts cytotoxic effects on a wide range of tumor cells and certain other target cells. Human TNF-A is a 17.4 kDa protein containing 157 amino acid residues.

Catalog#: RDI-301
|
Package Size: 50 micrograms $235.00    $219.00/package 50ug  2 or more

                    cat#RDI-301P Mini-Pack $95/10ug

                    SEE BULK BELOW

Supplied: Lyophilized powder from 3mM Tris pH 8.5 with no additives.

Source: E. Coli

Purity: >98% by SDS-Page analysis and HPLC analysis. Endotoxin level is less than 0.1ng per ug of TNF-Alpha.

Reconstitution: For best activity, reconstitute in 5mM Tris pH 8.0 to a concentration of   1mg/ml. (Can also be reconstituted in DI water)Allow to set at least 2 Hours at Temp, rotate, mix gentlyl. Centrifuge for 1-2 minutes at 1000 rpm to concentrate material in vial. This solution can be diluted into other buffered solutions containing a carrier protein or medium containing serum. and stored at 4 DEG C for up to 1 week or at -20 Deg C for future use. For most in-vitro applications, TNF-A exerts its biological activity in the range of 0.05 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.

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

Reactivity: This product is fully biologically active when compared to standards. The ED50 as determined by the cytolysis of murine L929 cells in the presence of Actinomycin-D is less than 0.05 ng/ml, which is a specific activity of > 2 X 10 E07 units/milligram.

AA Sequence:

          MVRSSSRTPS    DKPVAHVVAN   PQAEGQLQWL NRRANALLAN   GVELRDNQLV

           VPSEGLYLIY    SQVLFKGQGC    PSTHVLLTHT    ISRIAVSYQT       KVNLLSAIKS PCQRETPEGA

           EAKPWYEPIY  LGGVFQLEKG    DRLSAEINRP    DYLDFAESGQ    VYFGIIAL

Precautions: For In vitro research Use Only. Not for use in or on humans or animals or for diagnostics.

-see also rec murine TNF-A, rec rat TNF-A, and rec rabbit TNF-A.


Recombinant Human Tunor Necrosis Factor Alpha   (rHuTNF-A) BULK

cat# RDI-301-223X     $2188.00     $1750/mg 2-4   Bulk quotes on request

Description: Recombinant Human TNF-Alpha produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 158 amino acids and having a molecular mass of 17483.77 Dalton. The rHuTNFA is purified by proprietary chromatographic techniques.

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

Source: E.Coli

Endotoxin: Less than 0.1 ng/µg (IEU/µg) of TNF-A.

Formulation: The protein was lyophilized after extensive dialysis against 5mM tris buffer pH8.0.

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-VAL-Arg-Ser-Ser.

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

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

Stability: Lyophilized rHuTNF-A although stable at room temperature for 3 weeks, should be stored desiccated below -18 C. Upon reconstitution rHuTNF-A 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: TNF-a is fully biologically active when compared to standard. The ED50 as determined by the cytolysis of murine L929 cells in the presence of Actinomycin D is <0.05ng/ml, corresponidng to a Specific Activity of 2 X 10 E07 IU/mg

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

1. UV spectroscopy at 280 nm using the absorbency value of 1.234 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 TNF-a as a Reference Standard.

For In vitro research Use Only. Not for use in or on humans or animals .  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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