免疫组化求教

l****3

的两个病毒巨噬细胞炎症蛋白(vMIP)-I, -II, 我们有病理组织，已经通过PCR验证了病理组织中确实存在两个蛋白，现在要通过免疫组化来显示病理组织中存在的(vMIP)-I, -II，搜到文献一篇，但是本人因为没有做过免疫组化，所以对于文献提供的信息看不懂，下面有几个疑问帮忙解答一下

1.文章中没有直接提到vMIP)-I, -II抗体名字，是否两个蛋白的抗体可以直接找试剂公司找和买到。
2.本人也了解了一下免疫组化步骤，主要涉及到1抗和2抗，文中的2抗是否就是anti-rabbit IgG ，而我做的是人标本，是否可以用？
3，文中开始介绍了一段“Briefly, BC-3 cells (107 cells) in RPMI medium were induced with 20 ng/ml TPA (Sigma).

The cells were collected after 48 h of exposure to TPA, washed in phosphate-buffered saline

(PBS), pH 7.4, and spotted on slides in circles with 5-mm inner diameters, 24 circles per slide.”这段文字介绍的是什么？一般免疫组化准备好病理片子后，开始脱蜡水化，难道它说的步骤代替了脱蜡水化步骤吗？我们的组织都在病理蜡块里面。
4，哪位是这方面高手啊，可以直接指点我一下吗，我的QQ12607718,万分感谢。


Briefly, BC-3 cells (107 cells) in RPMI medium were induced with 20 ng/ml TPA (Sigma).

The cells were collected after 48 h of exposure to TPA, washed in phosphate-buffered saline

(PBS), pH 7.4, and spotted on slides in circles with 5-mm inner diameters, 24 circles per slide.

The slides were air-dried, then fixed in cold acetone for 10 min. The cells were then washed

with washing buffer (PBS supplemented with 0.1% Triton X-100) for 15 min, and incubated

with anti-vMIP-I or anti-vMIP-II monospecific antibodies (diluted 1:100 in IFA dilution

buffer [PBS containing 2% bovine serum albumin, 0.2% Tween-20, and 0.05% NaN2 ]) for

1 h at 37 ◦ C. After incubation, the slides were washed with washing buffer, then incubated

for 1 h at room temperature with a prestandardized dilution of rhodamine-conjugated goat

anti-rabbit IgG (Tago Immunologicals, Camarillo, CA). After washing, the slides were stained

with Hoechst 33342 to detect nuclei and mounted with 50% (v/v) glycerol in PBS. Three-

dimensional data sets were collected using a Deltavision microscope (Applied Precision,

Issaquah, WA), based on a Carl Zeiss inverted microscope with a Photometrics PXL cooled

CCD camera. The data sets were deconvolved, projected onto a single plane, and the contrast

was adjusted before the images were exported as TIFF files to Adobe Photoshop.