Polyclonal Goat and Monoclonal Mouse Anti-Rabbit Lg G - Essay Example

            In this graph, various dilutions of mouse anti-rabbit Ig G antibodies were used to determine the concentration of rabbit  Ig G. the most effective dilution will be between 1/4000 to 1/8000, as in the rest of the cases, at lower concentrations 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 linear relation. Similarly at more than 1/8000 dilution concentration of anti-rabbit IgG becomes too less compared to the concentration of rabbit Ig G. hence there will not be any anti-rabbit Ig G available to bind with rabbit Ig G at a higher concentration. In the given scenario the optimum dilution for mouse anti-rabbit IgG will be 1/6000 which gives a linear correlation with the increase in the concentration of rabbit IgG.

Graph 3: Polyclonal Goat anti-rabbit Ig G:

       Here in the case of goat anti-rabbit Ig G, the overall binding and absorption profiles restrict its use for the estimation of rabbit IgG. There are two different scenarios 1) In the case of lower dilution (1/2000) there is lots of nonspecific binding resulting in very high absorption at lower concentrations 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 the nonspecificity of polyclonal antibody. Due to less binding, there is no significant increase in absorption even at higher concentrations of rabbit Ig G. In conclusion polyclonal Goat anti-rabbit Ig G is not very useful for the determination of the titer of rabbit Ig G due to its nonspecific binding and specific activity.

        Based on the above three graphs it can be concluded that;

• The calibration curve has to be performed again In triplicates to get statically significant data.
• There should be the use of a multichannel pipette to avoid any sample-to-sample variation
• Mouse monoclonal anti-rabbit Ig G is a better choice at a dilution of 1/6000 to determine rabbit Ig G titer up to 2000ng/ml
• Goat polyclonal anti-rabbit Ig G will not be a good choice for this estimation due to higher nonspecific binding and can be avoided.
• But at extremely low concentrations of rabbit IgG, (less than 300ng/ml) goat polyclonal anti-rabbit IgG can be employed by diluting it to (1/4000) as it gives significant absorption. But one can not 100% rely on these results.