转移贴-请教：流感病毒 免疫组化

rojjer

xuqz005 发表于19/11/2010 16:51

大家好，我最近打算用病毒感染小鼠，然后取肺组织做切片，进行免疫组化检测，因为没有做过这方面的研究，不知道一抗和二抗分别用什么，有做过的朋友可否指教一下，谢谢

rojjer

llzh777：
你可以查查文献，看看前人用的什么？

suenmomo：
Pulmonary histopathology
Lungs were perfused, inflated and fixed in 4% formaldehyde as previously described and sections (4 mm thick) were prepared and stained with hematoxylin and eosin. Airway inflammation of hematoxylin and eosin-stained lung sections was scored on a scale of 0 1, 2, 3, 4 or 5 (corresponding to none, very mild, mild, moderate, marked and severe inflammation, respectively) on randomized, blinded sections by three independent readers. Tissues were graded for peribronchiolar inflammation (around 3–5 small airways per section) and alveolitis in multiple random fields per section as described. Immunoperoxidase staining of paraffin-embedded lung sections was performed to examine the distribution of viral antigens. Sections were subjected to antigen retrieval by heat treatment (10mM citrate buffer pH 6.0) and endogenous peroxidase activity was quenched with 0.3% H2O2 in methanol. Sections were blocked for 1 h (protein block, DakoCytomation, Glostrup, Denmark) and incubated with polyclonal goat-anti influenza A antibody (AbD Serotec, UK). The goat Vectastain ABC kit (Vector Laboratories, Burlingame, CA, USA) was then used in conjunction with 3,3¢ diaminobenzidine (DAB) substrate (ImmPACT DAB, Vector Laboratories) according to manufactures instructions. Sections were counterstained with hematoxylin (Sigma-Aldrich).
这只是一篇文献中的方法，涉及到肺部病理切片的都有免疫组化，楼主留心下就是
另外楼主也可以在pubmed里搜索influenza+Pulmonary histopathology

rojjer

xuqz005：
非常好，谢谢楼上的同学啦  
请问可不可以那你提及的那段文字的出处告知呢？谢谢

rojjer

suenmomo：
希望实验顺利！


xuqz005：谢谢版主。我也希望能按时毕业

haze:

一抗是病毒单抗， 二抗是都是公司买的