Mouse splenocytes were unstimulated (left) or stimulated for 5 hours with Cell Stimulation Cocktail (plus protein transport inhibitors) (Product # 00-4975) (right), then intracellularly stained with Anti-Mouse CD4 eFluor® 450 (Product # 48-0041) and 1 µg of Anti-Mouse/Rat TNF alpha PE-Cyanine7. Total viable cells, as determined by Fixable Viability Dye eFluor® 780, were used for analysis.
|Tested species reactivity||Mouse, Rat|
|Host / Isotype||Armenian hamster / IgG|
|Storage buffer||PBS, pH 7.2, with 0.1% gelatin|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C, store in dark, DO NOT FREEZE!|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1 µg/test|
|Functional Assay (FN)||Assay-Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Description: The TN3-19.12 monoclonal antibody reacts with mouse, rat, and rabbit tumor necrosis factor-alpha (TNF alpha), but not with human TNF alpha. The TN3-19.12 antibody is a neutralizing antibody. Mouse TNF alpha is a 17 kDa factor produced by macrophages, monocytes, neutrophils, CD4+ T cells and NK cells. A 26 kDa form of TNF alpha is expressed as a membrane-bound molecule. TNF alpha is cytolytic and plays an important role in immune regulation. Dimers and trimers of TNF alpha have been observed.
Applications Reported: This TN3-19 antibody has been reported for use in intracellular staining followed by flow cytometric analysis, cytokine neutralization, and ELISA.
Applications Tested: This TN3-19 antibody is offered in 2 formats: - µg size: has been tested by intracellular staining and flow cytometric analysis of stimulated mouse splenocytes. This can be used at less than or equal to 1 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. - test size: has been pre-titrated and tested by intracellular staining and flow cytometric analysis of stimulated mouse splenocytes. This can be used at 5 µL (1 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.
Light sensitivity: This tandem dye is sensitive 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 cell sample + 100 µL 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: 775 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser
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 noncovalently linked units, but shows a single ~17 kDa band following SDS-PAGE under non-reducing conditions.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.