Tumor necrosis factor alpha (TNF-α or cachectin) is a member of a family of small biologically active proteins known as cytokines. TNF-α plays a key role in the innate inflammatory response in vertebrates. When macrophages and T cells detect an invading pathogen, they produce TNF-α, triggering local production of chemokines and additional cytokines, which then attract and activate leukocytes at the infection site. (1,2,3,4) TNF-α also participates in the development of acquired immunity from the infection. (5) TNF-α plays a role in numerous other processes throughout the body, including remodeling of tissues and causing apoptosis of tumor cells. (6) Dysregulation of TNF-α production is involved in periodontal disease, heart disease, auto-immune conditions, and osteoarthritis. (7,8,9,10,11) Messenger RNA for TNF-α has been found in tissues of healthy human labial salivary glands, and the expression levels of the mRNAs were either up-or down-regulated by adjacent focal infiltrating lymphoid cells. (12)
The epithelial cells in the salivary glands are active participants in the autoimmune-mediated process of Sjögren’s syndrome, and TNF-α levels are increased in that disease. (12,13) Relationships between TNF-α levels in blood and saliva are not fully understood.
The effects of freeze thaw on most biological measures, regardless of biospecimen type, can be dramatic. Analytes in oral fluid are not distinct or different in this way. As a general rule, multiple freeze-thaws should be avoided. The most practical way to address this concern is by aliqouting samples after collection. Some analytes are more resistant to freeze thaw than others. We recommend that investigators consult the literature for the analytes of interest. If there is freeze thaw data for a specific biological measure in traditional biospecimens, it is reasonable to assume this would also be true for saliva.
No, but the literature is rather extensive on this subject for several salivary analytes; especially for salivary alpha-amylase and cortisol. We do not track that information internally.
With modern search tools online, we no longer maintain records of this type. We suggest that you use Pubmed or Psychlit to search the literature or you can Ask An Expert and we will be happy to assist you in your search.
If the analyte you are interested in is not noted in our website, please contact Dr. Granger at [email protected] to find out what developments are in the pipeline.
Salimetrics does not release products for sale if the quality control (QC) testing shows any issues. Here are some probable causes:
1) Can timing of adding reagents be off? For instance with a multichannel you can pipette the conjugate and TMB so many times before you refill, but you can pipette the stop twice as fast because it is a smaller volume. If you do this, it shortens the time the bottom rows have with TMB.
2) Can your washer be uneven and sheering off antibody in the bottom corner? Aspirate and check the amount of fluid left. It should be even in all wells and no wells should be completely dry.
3) Are you mixing faster than recommended? Or slower?
4) Are all reagents completely at room temperature? A bottle of assay diluent takes 2 hours to come to RT. You can pour some off into a smaller tube to warm up quicker for the zero and nsb.
5) Are you leaving the plate come to room temperature BEFORE opening the bag? (Otherwise moisture due to humidity may form in the wells and this is particularly a problem in this high humidity weather)
6) Are your multichannel pipettes dispensing the same amount each time reliably? We discard the first and last dispenses as they are not as reliable.
7) Are you incubating with TMB in the dark? (We no longer recommend aluminum foil.)
8) Are you testing one plate at a time? For example, do not put the standards on 5 plates then go back and fill in with samples. This delays the addition of conjugate to the plates.
9) Clean your plate reader filter. Dust from the lab can collect on the filter.
10) Are you adding assay diluent to the zero in sequence after the standards and not the last thing?
11) Never put the multichannel pipette tips into the wells as you can drag down standard from the wells above it causing lower readings in other wells.
12) Thoroughly blot all wells just before adding TMB but do not let the plate dry out.