Cell Bio Lab Report Essay

The purpose of this lab was to test the biological activity of ConA by performing a hemagglutinating activity assay. If ConA is active accordingly agglutination will occur due to ConAs free receptors being able to bind to the glucose residues on the sheeps red blood cells. If ConA is not active then no agglutination will occur. To test the hemagglutination reaction, two types of ConA antecedents were compared, a purchased keep back ConA solution in wing as the positive control, and a purified solution of ConA in buffer previously purified in lab. Each solution was at a 2mg/ml concentration of ConA in ConA buffer, which is necessary for ConAs biological activity. devil variables were added, Galactose and Mannose, to the ConA solution to compare the effects each had on the hemagglutination reaction. I hypothesize for ConA to be able to agglutinate the red blood cells if in the adequate concentration and if in the presence of Galactose, not Mannose. Mannose will inhibit the ConA fr om binding to the red blood cells membrane, preventing agglutination. RESULTSThe reaction plate containing the ConA dilutions was incubated all over the weekend and resulted in all wells being pink, appearing as if every well had agglutinated. There was a vague outline of the non-agglutinated cells in dissimilar wells. The last agglutination was observed at titer 0.0625 (1/16). Agglutination was seen in wordss A, B D, and E (row A contained the control ConA, row B contained the control ConA + Galactose, row D contained the sample ConA, and row E contained the sample ConA + Galactose). In the well rows C and F which contained control ConA + Mannose and sample ConA + Mannose, agglutination did not occur at any concentration of ConA. Row G, the negative control appeared to deem agglutinated as well as Row H, which contained only ConA buffer.DISCUSSION AND CONCLUSIONThe results did not tolerate my hypothesis for the biological activity of ConA. There are some sources of error that could explain the results obtained. Its possible there was a problem with either the ConA buffer or the sheep red blood cells to allow for all wells to turn pink and appear agglutinated. Another explanation of the irregular results was there might have been cross contamination from not changing tips when transferring to different ConA concentrations, or if bubbles were introduced while diluting the ConA, making the results difficult to interpret.For wells A, B D, and E as ConA became more thin out or decreased in concentration, it became more difficult for it to effectively crosslink and agglutinate the red blood cells. Well D, the positive control that contained the purchased ConA resulted in agglutination of the beginning(a) couple wells, then no agglutination as the ConA concentration decreased, similar to Row A. Wells B and E that had the Galacatose additive obtained the same titer of the control ConA because ConA does not bind Galactose. Galactose doesnt interfere with ConA f rom binding to the sugar residues on the red blood cells. Mannose on the other hand, is an inhibitor to ConAs binding sites. The Mannose in solution competed with the ConA and did not allow to bind to the sugar residues on the red blood cells as seen in rows C and F. Row G, the negative control, should have resulted in non-agglutination, similar to the rows containing the Mannose additive. The results observed delegateed agglutination formed in this row. Lastly, Row H should have shown non-agglutination through out because the well contained only ConA buffer, not ConA protein.In conclusion, the results did not clearly explain the biological activity of ConA with the hemagglutination assay. The experiment contained too many anomalies to get a clear determination of ConAs functionality post purification. The results did show that a change in the concentration of ConA would alter the strength of the reaction. Also, ConAs ability to bind to sugar residues can be affected if ConA has to compete or is inhibited to bind to a cells membrane. LITERATURE CITEDCell Biology 3822 Lab Manual, Cell Surface Glycoprotein Receptor AnalysisUsing Concanavalin A Lab 7. Pearson information Solutions. 2012 147-154.Madeleine Zaechringer. Cell Biology 3822 Analysis of purified ConA via Hemagglutinatino Assay Lab 7 Powerpoint. 2014.