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Human TNF-alpha Recombinant (CHO cell ),Biotinylated

TNFA

accession:  P01375
   Size A     10ug   $ 340
   Size B      25ug  $590
   Size C      100ug  $ 1950
Domain  :  TNF
Gene  :  TNFA
Catalog no. :  RBC01375
Source:
Optimized DNA sequence encoding Human Tumor Necrosis Factor-alpha  mature chain was expressed in CHO Cells.

 
Molecular weight:

Native human Tumor Necrosis Factor-alpha is generated by the proteolytic removal of the signal peptide and propeptide, this molecule has a calculated mass of approximately  18kDa.

Recombinant TNF-alpha is a disulfide-linked trimeric  protein, consisting of 3x158 amino acids and migrates as an approximately 18 kDa protein under reducing conditions.


 
Purity:
>95%, as determined by SDS-PAGE and HPLC

 

Biological Activity:

 

 

ELISA,WB

Biotinylated TNF-alpha was designed to be used in single step detection in ELISA ,WB.

The activity was measured by ability to bind immobilized TNFR1 in a functional ELISA. The ED50 of Biotinylated human TNFa is 0.5 μg/mL.


 
Protein Sequence:
        10         20         30         40         50         60 
MSTESMIRDV ELAEEALPKK TGGPQGSRRC LFLSLFSFLI VAGATTLFCL LHFGVIGPQR

        70         80         90        100        110        120 
EEFPRDLSLI SPLAQAVRSS SRTPSDKPVA HVVANPQAEG QLQWLNRRAN ALLANGVELR 

       130        140        150        160        170        180 
DNQLVVPSEG LYLIYSQVLF KGQGCPSTHV LLTHTISRIA VSYQTKVNLL SAIKSPCQRE 

       190        200        210        220        230 
TPEGAEAKPW YEPIYLGGVF QLEKGDRLSA EINRPDYLDF AESGQVYFGI IAL 
(*)Complete precursor sequence shown, expressed chain highlighted
 
Endotoxin:
Endotoxin content was assayed using a LAL gel clot method.
Endotoxin level was found to be less than 0.1 ng/µg(1EU/µg).

 
Presentation:
TNF-a was lyophilized from a 0.2μm filtered concentrated (1mg/ml) solution in PBS, pH 7.2.
Reconstitution:
A quick spin of the vial followed by reconstitution in distilled water to a concentration not less than 0.1 mg/mL. This solution can then be diluted into other buffers.

 
Storage:
The lyophilized protein is stable for at least 2 years from date of receipt at -20° C.
Upon reconstitution, this cytokine can be stored in working aliquots at 2° - 8° C for one month, or at -20° C for six months, with a carrier protein without detectable loss of activity.

Avoid repeated freeze/thaw cycles.

 
Usage:
This cytokine product is for research purposes only.It may not be used for therapeutics or diagnostic purposes.

 

TNF

Tumor Necrosis Factor is secreted by macrophages, monocytes, neutrophils, T-cells, NK- cells following their stimulation by bacterial lipopolysaccharides. Cells expressing CD4 secrete TNF- alpha while CD8(+) cells secrete little or no TNF- alpha. Stimulated peripheral neutrophilic granulocytes but also unstimulated cells and also a number of transformed cell lines, astrocytes, microglial cells, smooth muscle cells, and fibroblasts also secrete TNF. Human milk also contains this factor. The synthesis of TNF-alpha is induced by many different stimuli including interferons, IL2, GM-CSF, SP, Bradykinin, Immune complexes, inhibitors of cyclooxygenase and PAF (platelet activating factor). Human TNF-alpha is a non- glycosylated protein of 17 kDa and a length of 157 amino acids. Murine TNF- alpha is N-glycosylated. Homology with TNF-beta is approximately 30%. TNF- alpha forms dimers and trimers.

Related Publications:
tak1-mediated serine/threonine phosphorylation of epidermal growth factor receptor via p38/extracellular signal-regulated kinase: nf-b-independent survival pathways in tumor necrosis factor alpha signaling
Mol. Cell. Biol., Oct 2009; 29: 5529 - 5539.
tumor necrosis factor--mediated regulation of the inositol 1,4,5-trisphosphate receptor promoter
J. Biol. Chem., Oct 2009; 284: 27557 - 27566.
trovafloxacin enhances TNF-induced inflammatory stress and cell death signaling and reduces TNF clearance in a murine model of idiosyncratic hepatotoxicity
Toxicol. Sci., Oct 2009; 111: 288 - 301.
TNF- inhibition attenuates adverse myocardial remodeling in a rat model of volume overload
Am J Physiol Heart Circ Physiol, Oct 2009; 297: H1462 - H1468.
TNF inhibits apoptotic cell clearance in the lung, exacerbating acute inflammation
Am J Physiol Lung Cell Mol Physiol, Oct 2009; 297: L586 - L595.




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