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ELISA ASSAY - Essay Example

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The calibration curve is found to be good in terms of its statistical significance, as having R2 value of 0.94.And can be used as standard curve for Rabbit Ig G. There is slightly deflection at concentration of 2.0 this might be due to pipetting error as points before and after of it follows in line…
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ELISA ASSAY
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Download file to see previous pages In this graph various dilution of mouse anti rabbit Ig G antibodies were used to determined concentration of rabbit Ig G. the most effective dilution will be between 1/4000 to 1/8000, as in rest of the cases, at lower concentration of Ig G there is no linearity with increasing concentration of rabbit IgG (like in case of 1/2000). At1/2000 dilution concentration of mouse anti-rabbit Ig G is too high compared to rabbit IgG and hence there is no linier relation. Similarly at more than 1/8000 dilution concentration of anti rabbit IgG becomes to less compared to concentration of rabbit Ig G. hence there will not be any anti rabbit Ig G available to bind with rabbit Ig G at higher concentration. In given scenario the optimum dilution for mouse anti- rabbit IgG will be 1/6000 which gives linear correlation with increase in concentration of rabbit IgG.
Here in case of goat anti rabbit Ig G the over all binding and absorption profiles restricts it use for estimation of rabbit IgG. There are two different scenario 1) In case of lower dilution (1/2000) there is lots of non specific binding resulted in to very high absorption at lower concentration of rabbit Ig G and get saturated at slightly higher concentrations of rabbit Ig G.
2) In case of higher dilution (1/4000 and above) there is very less binding due to non specificity of polyclonal anti body. ...Download file to see next pagesRead More
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