Anti-Alpha Fodrin ELISA Kit
INTENDED USE
Human anti-alpha fodrin ELISA kit can be used for measuring in vitro quantitative levels of IgA or IgG class anti-alpha fodrin (anti alpha fodrin) autoantibodies in human plasma and serum samples. This assay has a minimum analytical sensitivity limit of 1.0 U/ml.
BACKGROUND
Alpha-fodrin is a dimer which is made up of an alpha- and a beta-subunit, it is an intracellular actin binding and a cytoskeleton organ specific protein. The dimer is cleaved to form a 120 kDa product during apoptosis. This 120 kDa product can be abundantly found in the salivary gland and is a vital autoantigen in pathogenesis. The heterodimers become part of the cytoskeleton structure where they are attached to the plasma membrane and can bind actin, microtubules and calmodulin.
Alpha fodin belongs to family of proteins that are known as spectrin, these are filamentous cytoskeletal proteins that are universally expressed within the body. They function in the organisation of organelles, secretion and molecular traffic. It is also one of the predominant targets for caspases during the processes of apoptosis, it is responsible for cell shrinkage and membrane malfunctions.
There is evidence to indicate that any abnormalities to alpha or beta-spectrin have been found to be linked to hereditary elliptocytosis. This is essentially a membrane protein spectrin structural change and can lead to many clinical manifestations such as RBCs defects. Clinical studies on patients with Sjögren Syndrome have shown that alpha-fodrin is also involved in the stimulation of peripheral blood T-cells. These findings indicated that presence of elevated levels of protease activity is responsible for T-cell stimulation and this activity is vital during primary Sjögren’s Syndrome development.
CONTENT
All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.
- Divisible Microplate: Coated with alpha-fodrin.
- Calibrators (A-F): Concentrations 0, 6.3, 12.5, 25, 50, 100 U/ml.
- Control Positive And Control Negative: Positive for IgG (1), positive for IgA (2) and negative for IgG and IgA (3).
- Sample buffer (5x).
- Enzyme Conjugate Solution: Polyclonal rabbit anti-human IgA and anti-human IgG; labelled with horseradish peroxidase.
- TMB Substrate Solution.
- Stop Solution.
- Wash solution (50x).
- Instruction for Use.
- Certificate of Analysis.
SENSITIVITY
The minimum detection sensitivity level of human IgG or IgA class anti alpha fodrin autoantibodies using current anti-alpha fodrin ELISA kit was 1.0 U/ml. The dynamic range for this assay is 6.3 – 100.0 U/ml.
ASSAY CHARACTERISTICS
– Results Interpretation: Negative: < 10 U/ml, Positive ≥ 10 U/ml
– Intra Assay Precision: 2.2 – 4.0%
– Inter Assay Precision: 1.1 – 3.4%
– Sensitivity: 1.0 U/ml.
– Specificity: Recognizes only autoantibodies specific to the alpha-fodrin.
– Interfering Substances: No interference has been observed with haemolytic (up to 1000 mg/dL), lipemic (up to 3 g/dL triglycerides) or bilirubin (up to 40 mg/dL) containing sera.
REFERENCES
- Sensitivity and specificity for primary Sjögren’s syndrome of IgA and IgG anti-alpha-fodrin antibodies detected by ELISA. J Rheumatol. (2004) 31 (3): 504-7. Ruffatti A., et al.
- Antifodrin antibodies in Sjogren’s syndrome: a review. Ann N Y Acad Sci. (2005) 1051: 235-9. Review. Witte T.
- The accuracy of the anti-α-fodrin antibody test for diagnosis of Sjögren’s syndrome: a meta-analysis. Clin Biochem. (2013) 46 (15): 1372-6. Review. Hu Q., et al.
- Molecular analysis of the human autoantibody response to alpha-fodrin in Sjögren’s syndrome reveals novel apoptosis-induced specificity. Am J Pathol. (2004) 165 (1): 53-61. Maruyama T., et al.
ADDITIONAL INFORMATION
- Full Name: Anti-Alpha Fodrin ELISA Kit
- Reactivity: Human
- Sample Type: Serum, Plasma
- Sensitivity: 1.0 U/ml
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