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Human anti-Tat IgG Immunoassay

Enzyme linked immunosorbent assay (ELISA) for the detection of anti-tat IgG in human serum and plasma, as an aid in monitoring the progression of disease in HIV-1 infected patients. Specifically, it could be useful for the screening and control of eligible patients in vaccination programs of HIV-1 infections with tat.


The HIV-1 regulatory tat proteins are considered attractive targets for the development of a multicomponent vaccine against HIV-1 infection. The protein is well conserved among different isolates and thus may be less susceptible to mutation leading to the production of escape virus variants. Tat is produced early after infection and it is essential for virus replication and infectivity. Tat protein is also immunogenic and antibodies against tat have been found to correlate with delayed disease progression and may exert protective effects by inhibiting both HIV replication and the effects of extracellular tat.


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Info

Microtiter strips coated with tat protein are incubated with collected samples. During this incubation step, anti-tat antibodies bind to the antigen forming specific complexes. Antibody excess is removed by washing and the antigen-antibody complex in each well is detected by adding an anti-human IgG HRP-conjugated antibody. Detection is performed by incubating the strips with 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) as chromogen and reading the absorbance values with a microplate reader at 415nm.

The kits allows 96 reactions; duplicates for each sample or control are strongly recommended.

Limitation of the Procedure:

• No drugs have been investigated for assay interference

• Do not use reagents beyond expiration date printed on the kit label

• Any variation in specimen diluent, operator, pipetting technique, washing technique, incubation time or temperature can cause variation in binding.

 

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