Description: The MP6-XT22 antibody reacts with mouse tumor necrosis factor-alpha (TNF alpha), a 17 kDa cytokine produced by monocytes, macrophages, neutrophils, NK cells and CD4(+) T cells. TNF alpha has cytolytic activity against a range of tumor cells and is important in immune regulation. TNF alpha forms dimers and trimers and also exists as a 26 kDa membrane-bound form.
Applications Reported: This MP6-XT22 antibody has been reported for use in intracellular staining followed by flow cytometric analysis.
Applications Tested: This MP6-XT22 antibody has been tested by intracellular staining and flow cytometric analysis of stimulated mouse splenocytes using the Intracellular Fixation & Permeabilization Buffer Set (cat. 88-8824) and protocol. Please refer to Best Protocols: Protocol A: Two step protocol for (cytoplasmic) iintracellular proteins located under the Resources Tab online. This can be used at less than or equal to 0.25 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
PE-eFluor® 610 can be excited with laser lines from 488-561 nm and emits at 607 nm. We recommend using a 610/20 band pass filter (equivalent to PE-Texas Red®). Please make sure that your instrument is capable of detecting this fluorochome.
Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.
Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.
Excitation: 488-561 nm; Emission: 607 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.
Filtration: 0.2 µm post-manufacturing filtered.
TNF alpha is a multifunctional proinflammatory cytokine that belongs to the tumor necrosis factor (TNF) superfamily. This cytokine is mainly secreted by macrophage and bind to its receptors, TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. TNF alpha is involved in the regulation of immune cells, cell proliferation, differentiation, apoptosis, lipid metabolism, and coagulation. TNF alpha exists as a multimer of two, three, or five noncovalently linked units, but shows a single 17 kDa band following SDS-PAGE under non-reducing conditions. Knockout studies in mice also suggested the neuroprotective function of TNF alpha, and has been observed to causes tumor necrosis when injected into tumor-bearing mice. Other functions of TNF-alpha include its role in the immune response to bacterial, viral, parasitic and certain fungal infections, as well as its role in the necrosis of specific tumors. TNF alpha causes cytolysis or cytostasis of certain transformed cells, being synergistic with interferon-gamma in its cytotoxicity. This cytokine has been implicated in a variety of diseases, including autoimmune diseases, insulin resistance, and cancer.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Protein Aliases: C-domain 1; C-domain 2; Cachectin; DADB-70P7.1; ICD1; ICD2; Intracellular domain 1; Intracellular domain 2; N-terminal fragment; NTF; TNF alpha; TNF superfamily; TNF α; TNF-a; TNF-alpha; TNFα; Tumor necrosis factor; Tumor necrosis factor ligand superfamily member 2; Tumor necrosis factor, membrane form; Tumor necrosis factor, soluble form; tumor necrosis factor-alpha
Gene Aliases: DIF; Tnf; TNF-a; TNF-alpha; Tnfa; TNFalpha; Tnfsf1a; TNFSF2
UniProt ID: (Mouse) P06804
Entrez Gene ID: (Mouse) 21926