This lab report describes an ELISA test conducted to determine immunoglobulin levels in samples. The ELISA test involves attaching antigens from samples to a plate and adding specific antibodies linked to enzymes. A detectable color change indicates the amount of antigens present. The students calculated immunoglobulin G levels in unknown samples by plotting absorbance readings against known concentrations to generate a standard curve equation. They then used the equation to determine the concentrations in the unknown samples were 27.5, 10.1, and -11.5 mg/ml, indicating hyper, normal, and no immunoglobulin G levels respectively.
This lab report describes an ELISA test conducted to determine immunoglobulin levels in samples. The ELISA test involves attaching antigens from samples to a plate and adding specific antibodies linked to enzymes. A detectable color change indicates the amount of antigens present. The students calculated immunoglobulin G levels in unknown samples by plotting absorbance readings against known concentrations to generate a standard curve equation. They then used the equation to determine the concentrations in the unknown samples were 27.5, 10.1, and -11.5 mg/ml, indicating hyper, normal, and no immunoglobulin G levels respectively.
This lab report describes an ELISA test conducted to determine immunoglobulin levels in samples. The ELISA test involves attaching antigens from samples to a plate and adding specific antibodies linked to enzymes. A detectable color change indicates the amount of antigens present. The students calculated immunoglobulin G levels in unknown samples by plotting absorbance readings against known concentrations to generate a standard curve equation. They then used the equation to determine the concentrations in the unknown samples were 27.5, 10.1, and -11.5 mg/ml, indicating hyper, normal, and no immunoglobulin G levels respectively.
Introduction: In this lab, we conducted an Enzyme Linked Immunosorbent Assay (ELISA) to determine immunoglobulin levels. Commonly, the ELISA test has been used as a diagnostic tool in medicine and pathology. The process is based on specific binding of an antigen via specific antibodies. Antigens from a sample are attached to a surface (i.e. well/microtiter plates) Then, a specific antibody is applied to bind to the antigen. This antibody is linked to an enzyme, and lastly a substance containing the substrate is added to the wells. The reaction produces a detectable colar change in the substrate. The ELISA test has many applications, such as identification and quantification of substances in tissues and body liquids, specifically hormones, drugs, markers, etc. Additionally, ELISA can be utilized to quantify antibodies in body tissues and fluids. In this lab, we calculated the HIgG from the sample patient and the primary antibody was linked to the HRP enzyme.
Lab Objective:
Calculations: After averaging the values for rows one and two and subtrating the average of the blank plates we came up with the following data
We plotted the absorbance vs concentration (Log). We then took the most linear data points in the center and had excel calculate our equation.
y = 3.2314x + 0.449
1.1361667 1.1831667 1.1906667 1.0926667 1.0126667 0.8386667 0.6361667 0.4446667 0.3206667 0.2441667 0.1831667 0.1726667 y = 3.2314x + 0.449 R = 0.9066 0 0.2 0.4 0.6 0.8 1 1.2 1.4 0.001 0.01 0.1 1 10 A b s o r b a n c e
( O D )
Concentration (mg/ml) Log The averages of our absorbance for the unkown samples are as follows as wella s the calculated concentrations: Unkown 1 Unknown 2 Unknown 3 1.338 .774 .076 Calculated concentration X 100 to correct for dilution 27.5 10.1 -11.5 Diagnosis Hyper HigG Normal HigG No HigG