产品展示
优质供应
详细内容
人血管紧张素Ⅳ(Ang-Ⅳ)elisa试剂盒
厦门仑昌硕生物科技有限公司 本公司集研发、生产、销售、服务及品牌代理为体的生物技术公司,目可提供各种抗体、免疫学相关的检测试剂盒、胎牛血清、动物抗血清等高品质产品及服务,涵盖分子生物学、细胞生物学、免疫学等生命科学域。价格实惠,质量有保证,请您放心选购,欢迎来电咨询!
Objective To explore the role and mechanism of non-receptor tyrosine kinase Tec in endotoxin/lipopolysaccharide (LPS)-induced production of interleukin-8(IL-8) in human alveolar epithelial cell A549. Methods Human alveolar epithelial cell A549 was cultured in RPMI-1640 medium containing 10% fetal bovine serum by volume for routine passage, and the second or third generation cells were taken for subsequent experiments. (1) The cells were taken according to random numbers. In LPS group, cells were cultured routinely for 1 hour, and then stimulated with 1μg/mL LPS for 1 hour. The cells in LFM-A13 group were treated with 75 μmol/L LFM-A13 in conventional culture medium for 1 hour, and then changed the culture medium for conventional culture for 1 hour. 25 μmol/L LFM-A13+ LPS group, 75 μmol/L LFM-A13+LPS group and 100 μmol/L LFM-A13+ LPS group were added with 25, 75 and 100 μ mol/l LFM-A13 in conventional culture medium for 1 hour. All the cells were changed and stimulated with 1 μg/mL LPS for 1 hour. The protein expression of Tec in each group was detected by western blotting, and the content of IL-8 in the supernatant of each group was detected by enzyme-linked immunosorbent assay. (2) The cells were randomly divided into 5 groups with 4 holes in each group. The cells in the blank control group were routinely cultured for 2 hours. The cells of small interfering RNA(siRNA) control +LPS group were transfected with empty lentivirus for 10 h, and then were stimulated by adding 1μg/mL LPS into conventional culture medium for 2 h. Tec mus-298 RNAi+LPS group, Tec mus-299 RNAi+ LPS group and Tec mus-300 RNAi+ LPS group were transfected with lentiviruses carrying tecmus-298 RNAi, tecmus-299 RNAi and tecmus-300 RNAi respectively for 10 hours. 1μg/mL LPS was added into the conventional culture medium to stimulate for 2 hours. the protein expression of Tec in each group was deTected by western blot method, and the Tec-siRNA.(3 with the best effect of silencing tec gene was screened. (3) the cells were divided into 4 groups according to random number table method, with 4 holes in each group. the cells in the blank control group were routinely cultured for 2 hours. The virus cells in the control group were transfected with empty lentivirus for 10 h, and then cultured routinely for 2h; The LPS group cells were stimulated by adding 1μg/mL LPS in the conventional culture medium for 2 hours. Tec-siRNA+ LPS group cells were transfected with lentivirus carrying tec-SiRNA with the best effect of silencing tec gene for 10 h, and then were stimulated by adding 1 μg/mL LPS in the conventional culture medium for 2 h. western blot was used to detect the protein expression of p38 mitogen-activated protein kinase (MAPK) and extracellular signal-regulated kinase (ERK)MAPK in each group. the data were analyzed by one-way ANOVA and LSD-T. 。
样品的处理及保存方法
人血管紧张素Ⅳ(Ang-Ⅳ)elisa试剂盒
1、 血清-----操作过程中避免任何细胞刺激。使用不含热原和内毒素的试管。收集血液后,1000×g离心10分钟将血清和红细胞迅速小心地分离。
2、 血浆-----EDTA、柠檬酸盐、肝素血浆可用于检测。1000×g离心30分钟去除颗粒。
3、 细胞上清液---1000×g离心10分钟除颗粒和聚合物。
4、 组织匀浆-----将组织加入适量生理盐水捣碎。1000×g离心10分钟,取上清液
5、 保存------如果样品不立即使用,应将其分成小部分-70 ℃保存,避免反复冷冻。尽可能的不要使用溶血或高血脂血。如果血清中大量颗粒,检测先离心或过滤。不要在37℃或更高的温度加热解冻。应在室温下解冻并确保样品均匀地充分解冻。
公司介绍
人血管紧张素Ⅳ(Ang-Ⅳ)elisa试剂盒
厦门仑昌硕生物科技有限公司 本公司集研发、生产、销售、服务及品牌代理为体的生物技术公司,目可提供各种抗体、免疫学相关的检测试剂盒、胎牛血清、动物抗血清等高品质产品及服务,涵盖分子生物学、细胞生物学、免疫学等生命科学域。公司非常注重企业信息化的建设及企业平台的建设,我们内部采用了的信息处理平台,保证客户的需求可以准确、高效处理。公司成立之初,立足外贸,和部分欧美及亚非国家的客户建立了良好的合作,随着国内生物域的蓬勃发展,逐步把重心转移到国内来,为每位科研人员献上高品质的产品及服务是我们的使命。
AMOY LUNCHANGSHUO BIOTECH CO.,LTD.