Experimental Procedures Of An Ige Antibody

In order to test for glucose, a glucose dip and read strip was placed in the first set of test tubes that were labeled bag start, solution start, bag end, and solution end. Then, a protein dip-and-read strip was placed in the same set

This experiment explores techniques of a titration, where a solution of a known concentration is used to determine the concentration of an unknown solution. Titrations are performed with a burette, which was created by Francois Antonie Henri Descrozilles, which contains the titrant, and a glass container such as a beaker or flask that contains the analyte.1 The titrant is solution of a known concentration that is slowly added to the analyte which is solution that is being analyzed to determine the unknown concentration of the solution. Inside the analyte is also an indicator that changes color near the equivalence point of titration. Titration have a large variety of uses, such as identifying the amount of binding proteins in the blood.2 The purpose of this experiment was to become familiarize and learn how to properly use a burette, pH probe, and pH indicator to effectively perform and examine a titration.

b. Attached to the secondary antibody is an enzyme such as peroxidase or alkaline phosphatase. These enzymes can metabolize colorless substrates (sometimes called chromagens) into colored products. After an incubation

This Lab Report is an analysis of the results of a two-part experiment. In the first part, we used a gel filtration column to separate the components of a mixture composed of protein and non-protein molecules. By doing so we hoped to obtain fractions that contained single components of the mixture, while also gaining insight into the relative molecular weight of each component compared to each other. We would then plot these fractions onto nitrocellulose paper in order to determine which fractions had protein. In the second part, we would use the fractions which we had determined had protein to conduct an SDS-PAGE. By doing so we hoped to determine an estimate on the molecular weight of the proteins present in each fraction by comparing it to a tracker dye composed of a variety of molecules of differing molecular weight.

Bradford assay would be ideal if an accurate estimation of a newly isolated protein using BSA as a standard. With the data obtain from the Bradford assay, it shows that it was more sensitive and quicker, also it had less interference from substance that were possibly present in the protein solution, expect for the interfering substance SDS.

Red and white blood cells are the two types of blood cells in the human body. Red blood cells transport oxygen around the body which is transferred through the bloodstream. It moves oxygen into the body and then removes it. They are absorbed through its haemoglobin.

Figure 1 contains gel electrophoresis for protein samples. The lanes were labeled from 1 to 10 from the right to the left. Lane 1 contained the ladder fragment. Lane 2 contained the filtrate. Lane 3 contained the S1 sample. Lane 4 contained the P1 sample. Lane 5 contained the P1 medium salt sample. Lane 6 contained the P1 high salt sample. Lane 7 contained the S2 sample. Lane 8 contained the P2 sample. Lane 9 contained the P2 medium salt sample. Lane 10 contained the P2 high salt sample.

Gel electrophoresis method. Qualitative analysis shows protein concentrations in kidney, heart, and liver. 1-6 are kidney tissue. 7-14 are liver and 15-19 are the heart tissue.

ELISA works on the principle of an antigen binding to specific antibody (lock and key), which can be used as a way to identify quantities of proteins in a small sample of fluid. The specific proteins used in an ELISA are estimated quantitatively. The ELISA test is carried out by incubating the serum that contains the antigen of interest with antibody’s within a well, in order for the antibody’s to bind with the specific antigens. The plate is then washed with a mild detergent in order to remove any proteins that have not been bound. The washing of the plates is carried out between every step in order

There are two methods using which the BCA Protein Assay can be performed- Test tube method and Micro plate method.

Ramos Vara JA, Miller MA. When Tissue antigens and antibodies get along: revisiting the technical aspects of immunohistochemsistry- the red, brown and blue technique. Vet Path. 2014; 51(1):42-87.

IgE antibodies live on the surface of cells that are specifically designed to be involved in the inflammatory response, mast cells and basophils. Mast cells are found predominantly in your surface tissues (tissues close to the external environment) such as in your skin and in the mucous membranes of your nose. Once the IgE has attached itself to an allergen, it sets off a series of events within the cells that lead to degranulation, which is the process that releases molecules from vesicles that regulate immune responses. Amongst these molecules is histamine.

3.2 The macrophages were plated at 18,000cells/cm2. Again to obtain the best result on the antibody and flow cytometric techniques, the cell concentration needs to be sufficient.

No lab tests were required at this visit, but if the physician would have decided for any blood work to support the diagnosis an immunoglobulin E(IgE) antibodies would have been appropriate to measure the immune system responds to the allergen milk. This test is not always conclusive in diagnosing cow’s milk allergy. CMP,CBC,C-reactive protein and stool exam could underline the presents of an inflammatory reaction. In addition, a skin test could be performed in order to confirm the presents of the milk allergy (Mayo Clinic, 2014).

The last part of this procedure, antibodies will be used to detect one specific protein from others on the membrane. Incubate the membrane with 10 ml of primary antibody for 10-20 minutes and then place on a rocking platform. Rinse the membrane quickly after with wash buffer on a rocking platform. After three minutes is up, discard the wash and incubate the membrane with 10 ml of secondary antibody for 5-15 minutes on the platform. Rinse the membrane again shortly after and wash the membrane for three minutes. Next, discard the wash and add 10 mL of HRP color detection reagent. Incubation will occur after this for 10-30 minutes. Once it is done, rinse the membrane twice with distilled water and blot dry.