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1、大鼠白介素1 (il-1)酶联免疫分析试剂盒使用说明书（ELISA kit for rat interleukin 1 beta (IL-1 beta)）ELISA kit for rat interleukin 1 beta (IL-1 beta)This kit is for research use onlyTest range: 3.12 pg/ml - 200 pg/mlMinimum detection limit: 0.78 pg/mlSpecificity: this kit allows simultaneous detection of natural or recombi
2、nant rat IL-1 beta and has no cross reactivity with other related proteins.Validity: 6 monthsProspective application: ELISA method for quantitative determination of IL-1, beta in serum, plasma, cell culture supernatant or other related biological fluids in rats.Explain1. kit kept: -20 degrees (when
3、not longer); 2-8 degrees (frequently used).2., concentrated liquid wash at low temperature will be salted out, diluted in the water bath heating, help dissolve.3., the English and Chinese manual may be out of order. Please use the English manual.4. the newly opened enzyme plate hole may contain a sm
4、all amount of water samples, this is normal, and will not have any effect on the results of the experiment.SummaryInterleukin -1 (IL-1), also known as lymphocyte stimulating factor, is divided into two kinds: IL-1 alpha and IL-1 beta. They are expressed by different genes, the molecular weight is 15
5、kDa, but the PI is 5 and 7 respectively. Both were composed of 159 and 153 amino acids, with only 26% homology between amino acid sequences, but were combined with the same membrane receptor. IL-1 is produced mainly by monocytes and macrophages in the blood. T lymphocytes, B lymphocytes, various epi
6、thelial cells, endothelial cells and stromal cells also produce IL-1. IL-1 in the blood is mainly IL-1 beta produced by monocytes and macrophages. IL-1 is a major regulator of acute immune response and plays a fundamental role in the activation of T cells and the induction of IL-2 production. These
7、regulatory actions are produced mainly by the action of the bone marrow stem cells in the nervous system. Most of these regulatory effects are directly produced by IL-1 beta, but some of these effects are synergistic with other cytokines such as IL-6, IFN, and TNF. Normal people do not contain IL-1,
8、 beta, or a very low level of blood. Elevated levels of IL-1 indicate tissue damage or infection in the body, such as stage enteritis, septicemia, etc. Inflammation and damage to the nervous system results in elevated levels of IL-1 in the cerebrospinal fluid. Elevated levels of IL-1 in the affected
9、 pleural exudate and peritoneal exudate.Experimental principlePurified antibody coated microtiter plate, made of solid support to the porous coated antibodies against IL-1 in specimens or standards, followed by adding biotinylated antibodies against IL-1, HRP labeled avidin, after thorough washing w
10、ith the substrate TMB color. TMB is converted to blue under the catalysis of peroxidase and converted to the final yellow under the action of acid. The color depth is positively related to the IL-1 beta in the sample. Measured absorbance in the 450nm wavelength with ELISA (OD), sample concentration
11、calculation.Kit composition and reagent preparation1. enzyme plates (Assay, plate): a piece (96 holes).2. standard (Standard): 2 bottles (freeze-dried).3. sample dilution (Sample, Diluent): 1 * 20ml/ bottles.4. biotin labeled antibody dilution (Biotin-antibody, Diluent): 1 * 10ml/ bottles.5. horsera
12、dish peroxidase labeled avidin dilution (HRP-avidin, Diluent): 1 * 10ml/ bottles.6. biotin labeled antibody (Biotin-antibody): 1 * 120 l/ bottles (1:100)7. horseradish peroxidase labeled avidin (HRP-avidin): 1 * 120 l/ bottles (1:100)8. substrate solution (TMB, Substrate): 1 * 10ml/ bottle.9. Buffer
13、 (Wash): 1 * 20ml/ bottle, use each bottle diluted with distilled water 25 times.10. termination liquid (Stop, Solution): 1 * 10ml/ bottles (2N, H2SO4).Reagents and equipment that are not needed1. standard and standard microplate reader2. high speed centrifuge3. electrothermal incubator4. clean test
14、 tubes and Eppendof tubes5. series adjustable pipette and suction head, it is better to use multichannel pipettes at one time for more samples6. distilled water, volumetric flask, etcCollection and preservation of specimens1. serum: whole blood samples should be placed at room temperature for 2 hour
15、s or 4 degrees overnight, and then centrifuged for 20 minutes at 1000 x g, Torikami Kiyo can detect or keep specimens at -20 degrees C or -80 degrees centigrade, but should avoid repeated freezing and thawing.2. plasma: EDTA or heparin can be used as anticoagulant, after sampling, 30 minutes C 1000
16、x g centrifuge 15 minutes, or specimens placed at -20 degrees C or -80 degrees to preserve, but should avoid repeated freezing and thawing 2-8 minutes.3. cell culture supernatant or other biological specimens: 1000 x g centrifugation for 20 minutes, Torikami Kiyo can detect, or put specimens at -20 degrees C or -80 degrees to preserve, but shoul
