Anti-TIF1-γ ELISA Kit is a semi-quantitative enzyme-linked immunosorbent assay (ELISA) for the detection of anti transcriptional intermediary factor 1-gamma - (TIF1-γ) antibodies in human serum.
The Anti-TIF1-γ ELISA Kit is an ELISA that measures anti-TIF1-γ antibodies in human serum sample. Calibrators and samples are added to the Microwells coated with recombinant human TIF1-γ antigen. During sample incubation, anti-TIF1-γ antibodies will react with the immobilized antigen. After washing to remove any unbound serum proteins, horseradish peroxidase conjugated anti-human IgG is added and incubated (Conjugate incubation). After another washing step, the peroxidase substrate is added and incubated for an additional period of time (Substrate incubation). Acid solution is then added to each well to terminate the enzyme reaction and to stabilize the color development. The test result is determined photometrically by measuring the absorbance and plotting the results.
1) TIF1-γ Microwell strips48 wells Microwell strips (8 x 6 wells) coated with recombinant TIF1-γ antigen. The breakaway strips are packed in a strip holder and sealed in a ziplock pouch with a desiccant.2)Calibrator 1 (0 U/mL)One vial containing 1.5 mL of assay diluent containing 0.09% sodium azide. Ready to use, make no further dilution.3) Calibrator 2 (100 U/mL)One vial containing 1.5 mL of anti TIF1-γ positive antibody in assay diluent containing 0.09% sodium azide. Ready to use, make no further dilution.4) ConjugateOne vial containing 8 mL of horseradish peroxidase conjugated goat anti-human IgG antibody and ProClin 150. Ready to use, make no further dilution.5) Assay diluentOne vial containing 50 mL of PBS, containing Tween 20 and 0.09% sodium azide. Ready to use, make no further dilution.6) Wash buffer (20x)One vial containing 50 mL of PBS with Tween 20 concentrate (20x).7) SubstrateOne vial containing 20 mL of 3,3',5,5'-tetramethylbenzidine dihydrochloride/hydrogen peroxide (TMB/ H2O2). Ready to use, make no further dilution.8) Stop solutionOne vial containing 20 mL of 0.25 mol/L sulfuric acid. Ready to use, make no further dilution.9) High controlOne vial containing 1.5 mL of anti TIF1-γ positive antibody in assay diluent containing 0.09% sodium azide. Ready to use, make no further dilution.10) Low controlOne vial containing 1.5 mL of human serum in assay diluent containing 0.09% sodium azide. Ready to use, make no further dilution.Unopened kit components are stable until the labeled expiration date at 2-8°C.
１． Targoff IN, et al. A novel autoantibody to a 155-kd protein is associated with dermatomyositis. Arthritis Rheum. 2006;54(11):3682-3689.２． Targoff IN, et al. Autoantibodies to transcriptional intermediary factor 1-gamma (TIF1-g) in dermatomyositis [abstract]. Arthritis Rheum. 2006;54 Suppl:S518.３． Kaji K, et al. Identification of a novel autoantibody reactive with 155 and 140 kDa nuclear proteins in patients with dermatomyositis: an association with malignancy. Rheumatology (Oxford). 2007４． Fujimoto M, et al. Myositis-specific anti-155/140 autoantibodies target transcription intermediary factor 1 family proteins. Arthritis Rheum. 2012 Feb;64(2):513-22.５． Fujimoto M. [Myositis-specific autoantibodies]. Brain Nerve. 2013;65(4): 449-460