Mouse Anti-TNF-alpha (APC1, Cachectin, Differentiation inducing factor (DIF), Macrophage cytotoxic factor (MCF), Necrosin, TNF alpha, TNF Macrophage Derived, TNF Monocyte Derived, TNF Superfamily Member 2, TNFA, TNFSF2, Tumor necrosis factor ligand superfamily member 2, Tumor Necrosis Factor Precursor) (FITC)
(APC1, Cachectin, Differentiation inducing factor (DIF), Macrophage cytotoxic factor (MCF), Necrosin, TNF alpha, TNF Macrophage Derived, TNF Monocyte Derived, TNF Superfamily Member 2, TNFA, TNFSF2, Tumor necrosis factor ligand superfamily member 2, Tumor Necrosis Factor Precursor)
Tumor Necrosis Factor Alpha (TNF alpha) is a protein secreted by lipopolysaccharide-stimulated macrophages, and causes tumor necrosis when injected into tumor bearing mice. TNF alpha is believed to mediate pathogenic shock and tissue injury associated with endotoxemia. TNF alpha exists as a multimer of two, three, or five non-covalently linked units, but shows a single 17kDa band following SDS PAGE under non-reducing conditions. TNF alpha is closely related to the 25kDa protein Tumor Necrosis Factor beta (lymphotoxin), sharing the same receptors and cellular actions. TNF alpha causes cytolysis of certain transformed cells, being synergistic with interferon gamma in its cytotoxicity. Although it has little effect on many cultured normal human cells, TNF alpha appears to be directly toxic to vascular endothelial cells. Other actions of TNF alpha include stimulating growth of human fibroblasts and other cell lines, activating polymorphonuclear neutrophils and osteoclasts, and induction of interleukin 1, prostaglandin E2 and collagenase production. TNF alpha is currently being evaluated in treatment of certain cancers and AIDS Related Complex.
Applications: Suitable for use in Immunofluorescence, ELISA, Flow Cytometry, Immunoprecipitation, Immunohistochemistry. Other applications not tested.
Recommended Dilution
Flow Cytometry: 0.5-1ug/million cells Immunofluorescence: 1-2ug/ml Immunoprecipitation: 1-2ug/500ug protein Immunohistochemistry: Frozen & Formalin-paraffin: 1-2ug/ml for 30 min at RT (Staining of formalin-fixed tissues requires boiling tissue sections in 10mM Citrate Buffer, pH 6.0, for 10-20 min followed by cooling at RT for 20 minutes Optimal dilutions to be determined by the researcher.
Positive Control
HeLa, HL-60, or A431 cells. Macrophages in lymph node or Tonsil
Storage and Stability
Store product at 4°C if to be used immediately within two weeks. For long-term storage, aliquot to avoid repeated freezing and thawing and store at -20°C. Aliquots are stable at -20°C for 12 months after receipt. Dilute required amount only prior to immediate use. Further dilutions can be made in assay buffer. Caution: FITC conjugates are sensitive to light. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Note: Applications are based on unconjugated antibody.
Immunogen
A hexadecapeptide corresponding to aa115-130 of human TNF-alpha, conjugated to thyroglobulin. Cellular Localization: Cytoplasmic and extracellular (secreted)
Form
Supplied as a liquid in 10mM PBS. No preservative added. BSA-Free. Labeled with Fluorescein isothiocyanate (FITC).
Purity
Purified by Protein A/G affinity chromatography.
Specificity
Recognizes human TNF-alpha. Species Crossreactivity: mouse, rat, rabbit, feline, canine, and zebrafish. Others-not known.
