Test1. Production全面从严治党 of antibody
[principle]
1. In the special humoral immune respon, antigen can stimulate immune system to produce specific antibodies. Each antigen molecule has veral different antigenic determinants, so it can be recognized by different B cells and then the B cells will produce different specific antibodies that can bind with epitopes specifically. The immunity rum produced by using antigen to immune animal is a mixture of many different antibodies, called polyclonal antibody. The most common method to produce polyclonal antibody is using pure antigen to immune animal.
2. The factors influencing producing of antibodies is related to antigen purity, animal, amount of antigen, ways of injection, times of immunity, etc.
Primary immune animal, the antigen enter animal for the first time, the body will produce a small amount antibodies after a latency, but when cond injection antigen to the animal, th
ere will be a fast respon to the antigen and a lot antibodies will be produced. So in this test, u antigen to immune mice 3 times, then we can detect antibodies in the rum.
[methods]
Preparation of antigen:
1. add 3 ml N.S to tube containing SRBC and mix them.
2. centrifuge 2000rmp, 5 min.
3. 蓝色的寓意和象征discard the supernant
4. repeat above operation.
5. U N.S to dilute and make 20% 、40% SRBC solution.
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6. Take 0.1ml 40% to immunize the mice subcutaneously.小小怪兽
7. Do the condary and third injection after every week.
Test 2.ELISA
[principle]
1. Immunological labeling techniques: Ag-Ab reactions are combined with labeling techniques. Known Ag or Ab is labeled with radioisotope, enzyme or fluorescein and unknown Ab or Ag are indirectly detected by labeled molecules. It can divided into Radioimmunoassay(RIA), Immunofluorescence technique and Enzyme Immunoassay. The Direct labeling techniques are to label each Ag and Ab, the indirect labeling techniques are to label condary Ab.
2. Enzyme immunoAssay (EIA): Ag-Ab reactions with enzyme-labeled Ag or Ab. Usually using horradish peroxida (HRP)，alkaline phosphata (AP) to label Ag or Ab. It can be divided into ELISA and immunochemistry.
3. ELISA: Unknown Ab or Ag in blood or culture medium are detected by enzyme-labeled Ag or Ab. It can be divided into 3 methods, Indirect ELISA(Known Ag, enzyme-labeled condary Ab to detect unknown Ab ),Sandwich ELISA and Competitive ELISA.
First, the known antibody or antigen is fixed on a solid carrier. Then a sample and the corresponding enzyme-linked Ab/Ag are added to bind to the Ag/Ab attached to the solid carrier. The resulting Ag-Ab complex and the excess Ab/Ag are parated by washing. Finally the substrate is added and catalysis cau a color change which can be measured.ELISA has the advantages of high nsitivity and specificity. It combines the specificity of the Ab-Ag reaction with the catalysis of enzymes.
[methods]
1. Preparation of Ag: add 3ml N.S to tube containing defibrated SRBC and mix. Then centrifuge 2000rmp, 5 min. add 3ml N.S to the tube and mixed, then take 2 ml packed SRBC. Add 2ml DDW and shatter SRBC and dilute with coating buffer in a ratio of 1:400.电脑耳机没声音怎么设置
2. Coating Ag: Add 100μL of Ag to each well of ELISA plate, then u N.S to wash for five times.
3. Prepare the rum: remove the eyeball of the SRBC-immunized mice and collect the bl
ood into Ep tube. Wait for about 5 minute then centrifuge at 12000rmp for 10min. take the supernant and dilute the rum 1:10.
4. Add test rum: sign and add 100μL of solution to each well, then put it in a humidified box under 37℃ for 45min威斯特伐利亚和约.
5. Add condary Ab: Discard the solution in ELISA plate and wash the plate 3 times. Then add 100μL of HRP-labeled condary Ab to each well and put it in a humidified box under 37℃ for 30min.
6. Showing color: Discard the solution in ELISA plate and wash the plate 3 times. Then add 100μL of substrate solution to each well. Show color in dark for10min
[result]
the No.1 well is blank, No.2 is negative well, No.3 is positive well, No.4 is test well. The No.3 and No.4 became yellow while No.1 and No.2 is colorless. The color of the No.4 well is deeper than the No.3.绿色校园作文
[analysis]
The result in No.4 well is positive becau it become yellow,which means there are antibodies in the No.4 well also indicates that SRBC- immunized mice had produced antibodies. The color of test well (No.4 well) is deeper than the positive control well(No.3 well), so antibodies amount in test rum is more than that in the positive control.
Test3. 梦见自己来例假了Hypernsitivity Test of Guinea Pig
[principles]
Hypernsitivity refers to the lf tissue injury and/or biological disorder when the nsitized individual contacts the same allergen again. Hypernsitivity is the abnormal or pathologic immune respon. According to the mechanisms of hypernsitivity, it could be divided into four types, typeⅠ、Ⅱ、Ⅲ and Ⅳ. Type I hypernsitivity is also called immediate type hypernsitivity or anaphylaxis. The antigen leads to hypernsitivity is called allergen. The antibody take part in anaphylaxis is mainly IgE.
