The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application notesELISA: Use at a concentration of 1 µg/ml to detect 0.6 ng/well of rhTNF beta.
Neut: Use at a concentration of 0.1 - 1.6 µg/ml.
WB: Use at a concentration of 2 µg/ml to detect rhTNF beta at 5 ng/lane. Predicted molecular weight: 22 kDa.
Anti-Human TNF beta is tested for its ability to neutralize the biological activity of rhTNF beta on L929 cells. The ND50 of the antibody is defined as the concentration of antibody required for one-half maximal inhibition of bioactivity of rhTNF beta which is present at 5 times its own EC50 (the concentration of rhTNF beta producing a one-half maximal bioactivity without antibody). In this bioassay, rhTNF beta was pre-incubated with various dilutions of the antibody in culture medium containing actinomycin D for 1 hour at 37°C in a 96-well microtiter plate. Following this pre-incubation period, the antibody-antigen mixture was added to confluent cultures of L929 cells in 96-well microtiter plates to give a final concentration of 1 ng/ml rhTNF-beta and 1 ug/ml actinomycin D in 0.1 ml. This was incubated for 24 hours at 37°C in a 5% CO2 humidified incubator and then fixed with 5% formaldehyde and stained with crystal violet.
Not tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
FunctionCytokine that in its homotrimeric form binds to TNFRSF1A/TNFR1, TNFRSF1B/TNFBR and TNFRSF14/HVEM. In its heterotrimeric form with LTB binds to TNFRSF3/LTBR. Lymphotoxin is produced by lymphocytes and cytotoxic for a wide range of tumor cells in vitro and in vivo.
Involvement in diseaseGenetic variations in LTA are a cause of susceptibility psoriatic arthritis (PSORAS) [MIM:607507]. PSORAS is an inflammatory, seronegative arthritis associated with psoriasis. It is a heterogeneous disorder ranging from a mild, non-destructive disease to a severe, progressive, erosive arthropathy. Five types of psoriatic arthritis have been defined: asymmetrical oligoarthritis characterized by primary involvement of the small joints of the fingers or toes; asymmetrical arthritis which involves the joints of the extremities; symmetrical polyarthritis characterized by a rheumatoidlike pattern that can involve hands, wrists, ankles, and feet; arthritis mutilans, which is a rare but deforming and destructive condition; arthritis of the sacroiliac joints and spine (psoriatic spondylitis).
Sequence similaritiesBelongs to the tumor necrosis factor family.
Cellular localizationSecreted. Membrane. The homotrimer is secreted. The heterotrimer is membrane-associated.