Purpose In the Affinity Chromatography experiment we were purifying our Con A proteins. In general, affinity chromatography is a technique that is used for isolating a protein, in our case Con A from a large amount of other macromolecules. Our protein of interest is captured using a microbead matrix while we let everything else flow through the column. The Sephadex matrix is made of cross-linked glucose or dextran and because our Con A has an affinity for glucose it is able to bind to those beads. In general, we began by equilibrating our column with NaCl, then poured Jack Bean Meal Extract which so happens to contain Con A through our column, the Con A then binded to the Sephadex beads, and finally we eluded with a dextrose solution so that …show more content…
We began the experiment by mixing our Con A with sheep red blood cells. If Con A is biologically active it is able to agglutinate the red blood cells. If Con A is inactive it results in no agglutination. Partial agglutination is another possible result. Our experimental variables were Con A + mannose and Con A + galactose. Ultimately we were comparing our purified Con A sample activity to a controlled Con A …show more content…
We found that Tube 1 had an absorbance of 0.085, Tube 2 had 0.034, Tube 3 had 0.027, Tube 4 had 0.032, Tube 5 had 0.025, Tube 6 had 0.028, Tube 7 had 0.022, Tube 8 had 0.024, Tube 9 had 0.030, and Tube 10 had an absorbance of 0.022. We then graphed absorbance vs fraction number so that we could demonstrate the absorbance of each tube and found that Tube 1 had our highest value of absorbance. Basically, the first few tubes have the most protein and then the rest drop off as is shown in the graph. For the hemagglutination assay experiment the Con A concentration went from full strength at well A1 and at well A12 was the least. In the row of Con A by itself there was agglutination present in A1 through A6 but eventually the concentration became so dilute that around A7 the red blood cells were not able to agglutinate anymore. Similar results were seen with our Con A + galactose. Our Con A + mannose showed the first few wells with agglutination but eventually our Con A was not able to agglutinate the red blood cells due to the mannose interference. The negative control showed no agglutination throughout the wells. Our Con A wells should’ve been similar in comparison to the wells with control Con
The purpose of blood culture draw is to test for the existence of bacteria and other microorganisms that could possibly be in a patient’s blood. Even the smallest contamination will have major adverse effects on the final results of this kind of test. The second in the order of draw is the one use for a coagulation assay. The drawing tube that is use for a coagulation assay has a light blue top. When it comes to coagulation assay, there is an exemption to the order of draw. If it is believe that tissue fluids or thromboplastins have infected that blood, then a non-additive tube may be used in replace of the tube for the coagulation assay. The non-additive tube usually has a red top on it. The third tube in the order of draw is the non-additive, which I said earlier is marked with a red top. This tube is for blood draw taken for testing without adding any kind of reaction additive. The fourth and final draw in the order of draw is an additive tube. There are many different additives that can be used for testing purposes. The final measure of the order of draw has an order in itself that must be followed. The first tube used for additive is the one that contains SST. This tube usually has a red-gray or gold top on it. This tube has additives that have gel separators and clot activator. This tube is used for the testing of the pancreas function in the patient.
Blood is a non-Newtonian fluid that contains many components. One such component is red blood cells. Due to the red blood cells having the tendency to clump together at low velocities, calcium chloride was added in order to cause a thrombus formation. The blood used in this experiment was sheep’s blood.
The goal of this experiment is to determine the blood types of the samples given and to learn what interactions occurred to each blood type. Determining an individual’s blood type and how it reacts with Anti A, Anti-B, and Anti Rh serums played a crucial part in this experiment. The researcher concluded that agglutination (clumping) occurred in some of the blood samples. For example, Mr. Smith’s blood reacted with Anti-A and Anti-Rh serums (antibodies) allowing the researcher to determine the blood type is A. Mr. Jones’s blood reacted with Anti-B serum but it did not react to Anti-A or Anti Rh allowing the researcher to believe that the blood type is B. Mr. Green’s blood reacted with all serums and caused a reaction to occur resulting the blood type to be AB positive. Mr. Green’s blood also had a positive marker for Rh factor. However, Ms. Brown’s blood had no reaction at all and the researcher determined if no reaction occurred then the sample had no antigens but proved to have some antibodies, resulting in blood type to be O. The purpose of this experiment is to determine whose blood has type A, B, AB, or O.
Hypothesis – With the antigen and antibody reaction we can determine ABO and Rh blood types by using certain antiserums. If an A Antigen ever comes in contact with an Anti a antibody then the blood will have a reaction. Also if a B Antigen comes in contact with a B anti body then the blood will
In a cell, diffusion occurs across a selective permeable membrane of the cell that allows the cell to regulate and control the passage of substances. As well as controlling what passes through the membrane, the selective permeability also controls how quickly those allowed substances are able to pass through the cell membrane. When a cell is placed in a hypotonic solution, the substance wants to quickly diffuse through the cellular membrane in attempt to balance the concentration. Due to this, too much of the substance enters the cell causing the cell to swell and rupture, otherwise known as hemolysis. The purpose of this lab is to test if and how the factor of polarity of a molecule affects the diffusion rate of those molecules through a permeable red blood cell membrane in a sample of ovine blood. Testing this factor and its effect on diffusion rate is important in the biological world as it helps understand how our body regulates our own substances and molecules within us, but also how our body will react and respond to other substances such as medicines. It is hypothesized that this factor of polarity will determine how quickly or slowly molecules will diffuse across the cell membrane. Smaller molecules that have relatively low polarity will be able to cross the membrane more easily and at a faster rate than those molecules that
The specificity of albumin binding experiment was to determine the binding interactions that occur between serum albumin and three synthetic dyes with the use of electrophoretic procedure. Whole blood, or plasma. Clots upon standing and if the clot is removed, the remaining straw colored fluid is called serum. The major protein in serum is albumin which functions as a carrier molecule for the transport of certain small molecular weight compounds in blood. Molecules that bind to serum albumin are fatty acids, hormones and some synthetic dyes. In this experiment the synthetic dyes used are Bromophenol Blue, Ponceau S and Orange G. we observed that free dyes not bound to albumin migrate faster that albumin or dyes bound to albumin. This
Blood is a non-newtonian fluid that contains many components. One such component is red blood cells. Due to the red blood cells having the tendency to clump together at low velocities, calcium chloride was added in order to cause a thrombus formation. The blood used in their experiment was sheep’s blood.
In a labeled glass tube, two drops of the suspension are mixed with one drop each of commercially prepared anti-A, anti-B and anti-D. After mixing, the tubes are centrifuges at low speed in a serofuge, the tubes are gently rocked/shaken so that the fluid flows across the pelleted RBCs. The pellet is observed carefully for agglutination using a magnified mirror. The reverse typing is accomplished in the same manner using two drops of patient serum and one drop of commercially prepared A1 cells and B cells. Figure 3 shows the criteria used for macroscopic grading of agglutination
There are some errors that can happen while using a haematocrit in the lab. such as hemolyzed sample can give an incorrectly decreased to the results and it should not be used. The blood should be well-mixed and be placed at room temperature before testing. The capillary tube must be well sealed former to be the centrifuge. If it is not sealed, the result will be incorrectly low because when the tube is being centrifuged, red cells will be escaping throughout the gap in the seal. The blood anticoagulant ratio is very important, especially when using EDTA. Excessive EDTA can give a falsely decreased hematocrit due to the shrinkage of the red blood cells. The liquid anticoagulants, such as sodium citrate and heparin can give falsely decreased
When I stirred the first sample with each type of antiserum, the blood became cloudy and particles formed in it when the antisera-A and antisera-Rh were stirred in, so that showed a reaction. Because it reacted with only these two types of antiserum, that combination shows that the blood is AB Positive because of the combination of antibodies in the blood. AB Positive blood has both A and B antigens on the red blood cells, but neither A or B antibodies on the plasma. For the second sample, I stirred in all three serums and the blood reacted with Anti-B and Anti-Rh. Because it reacted with these two serums, it shows that the blood has the combination of antibodies that make it type B Positive. This means that there are B antigens on the red blood cells, but A and B antibodies are in the plasma. For the third sample, I stirred in all three types of serum, and it did not react with any of the serums. All three serums resulted in clear, red blood that was not cloudy with no particles formed. When this happens, it shows that the blood is O Negative because it did not react with any of the antibody-stimulating serums. This means that there are neither A or B antigens in the red blood cells, but both A or B antibodies in the plasma. For the fourth sample, I stirred all the serums in to the sample. It became cloudy when the Anti-A serum was stirred in, but the other two
After the dogs had been anesthetized and given the injection of the endotoxin, blood was drawn. The blood was collected by use of a catheter in the femoral artery. However, for each sample, a new catheter was used. Between each collection of the samples, the artery had to be clamped using a Pott’s Clamp. The blood collected was then stored in a silicon tube. Nine units of whole blood were anticoagulated with one volume of 3.2% sodium citrate. The plasma was then separated by centrifugation for 30 minutes and stored in silicon test tubes at negative twenty
The purpose of this experiment was to determine the effects of tonicity on a cell membrane using red blood cells, potato strips and three unknown solutions (A, B, C). First three slides were prepared containing RBC’s and unknown solutions A, B and C. A control slide was prepared only using RBC’s. After observing each slide under the microscope it was determined that unknown solution A was hypertonic because the RBC appeared to have shrunk. The RBC in unknown solution B appeared to be swollen, therefor, the tonicity of unknown solution B was hypotonic. Unknown solution C showed no change to the RBC shape, it was suggested that unknown solution C was isotonic. To confirm the tonicity
The goal is to describe the lytic process, using the activation of the alternative pathway. In the laboratory, sheep red blood cells are used and fish cells are introduced as foreign body cells. Complementary activities are investigated by recording the haemoglobin released from lysed erythrocytes.
Therefore, the hemolytic activity must be considered by oneself as an unreliable criterion to detect of biosurfactant activity of a microbial culture [53]. The hemolytic activity of biosurfactants was first discovered when Bernheimer and Avigad [7] reported that the biosurfactant produced by Bacillus subtilis, surfactin, lysed red blood cells.
2.The agglutination is based on binding of the sialic acid to the cell membrane. To confirm that sialic acid carbohyrates are responsible for the binding the cells can be stripped of sialic acid and the experiment repeated.