求助小鼠CD8免疫组化问题
免疫组化（石蜡）测小鼠肿瘤中CD8+CTL浸润情况，一抗为Biolengend的大鼠抗小鼠CD8a（克隆号：53-6.7），结果很不理想，换个一抗测巨噬细胞，其他所有试剂都一样，结果很好。
是CD8抗体的问题吗？请各位前辈帮忙分析。
有谁用过这个公司的抗体，或测过小鼠CD8，请前辈们帮忙。
我的染色步骤：(SP试剂盒、DAB均为中山的，二抗为羊抗大鼠IgG)
常规脱蜡至水－－水洗，3％H2O2阻断－－柠檬酸抗原修复－－封闭－－－一抗4℃过夜－－复温－－二抗－－过氧化物酶标记的链酶亲和素－－DAB显色－－苏木素复染－－分化－－脱水透明。
说明书：
Purified anti-mouse CD8a
Product Data Sheet
Catalog #Size: 100701 / 50 µg
100702 / 500 µg
Clone: 53-6.7
Isotype: Rat IgG2a, κ
Immunogen: Mouse thymus or spleen
Reactivity: Mouse
Preparation: The antibody was purified by affinity chromatography.
Formulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Storage: The antibody solution should be stored undiluted at 4°C.
Applications
Applications: FC, IHC(Frozen/ Paraffin), IP
Application：Additional reported applications (for the relevant formats）include:
Notes: immunoprecipitation, in vivo and in vitro cell depletion3, inhibition of CD8 T cell proliferation4, blocking
of cytotoxicity，and immunohistochemical staining of acetone-fixed frozen sections, zinc-fixed paraffin- embedded sections and 10% formalin-fixed paraffin-embedded sections. The 53-6.7 antibody
competes with 5H10-1 antibody (Cat. No. 100802) for binding to thymocytes4. The LEAF™
purified antibody (Endotoxin <0.1 EU/μg, Azide-Free,0.2 μm sterile-filtered) is recommended for
functional assays (Cat. No. 100716).
[ 本帖最后由 flowing_water 于 2007-7-5 10:44 编辑 ]
