说明书丨Fluorochorome荧光金(Fluoro-Gold)抗体

Fluoro-Gold can be injected using several different methods, including pressure, iontophoretic and otherapplications developed by a variety of researchers. See Schmued and Fallon, Fluoro-Gold: "A

fluorescent retrograde axonal tracer with numerous unique properties”, Brain Research, 377 (1986) 147- 154 as well as Pieribone and Aston-Jones, "The Iontophoretic Application of Fluoro-Gold for the study ofaf erents to deep brain nuclei”, Brain Research, 475 (1988) 259-271. Many researchers have developedtheir own modified procedures. Use of the antibody should not be dependent upon the methodology usedto employ Fluoro-Gold.

After the Fluoro-Gold has been injected, floating sections (we used thirty um sections from a rat perfusedwith 4% formaldehyde) are incubated with the Fluoro-Gold Antibody solution overnight at 4º C. Sectionsare washed, then incubated in Biotinylated GAR (Vector Labs) at 1/1000 for 1 hour at room temperatureand washed again. Sections are then incubated in Avidin/Biotin (Vector Labs) at 1/1000 for 1 hour, washed and transferred to Diaminobenzidine (.04%) and Nickel Chloride (2.5%) in 0.1 M NaAcetate with0.06% H2O2 for six minutes. Sections are then washed, mounted, dried, dehydrated and cover slipped.

It has been our experience that if stored and prepared in the manner set out above, each vial of the antibody should treat 300 to 1000 thirty um sections of the albino rat brain (or similar sized animals) ifyou use the Vector elite kit. However, you should experiment with the concentration and procedures todetermine which best fit your circumstance.

艾美捷Fluorochorome荧光金(Fluoro-Gold)抗体相关介绍：

氟金可以用几种不同的方法注射，包括压力、离子电渗和各种研究人员开发的治疗方法。参见Schmoud和Fallon，Fluoro Gold：“A荧光逆行轴突示踪剂具有许多独-特的财产”，《大脑研究》，377（1986）147-154，以及Pieribone和Aston-Jones，“荧光金在研究脑深部细胞核的遗传物质中的离子导入应用”，《脑研究》，475（1988）259-271。许多研究人员开发了他们自己的改进程序。抗体的使用不应依赖于使用荧光金的方法。

注射Fluoro Gold后，将漂浮切片（我们使用了灌注了4%甲醛的大鼠的30μm切片）与Fluoro金抗体溶液在4℃下孵育过夜。切片被洗涤，然后在室温下在生-物-素化GAR（Vector Labs）中以1/1000孵育1小时，然后再次洗涤。然后将切片在Avidin/Biotin（Vector Labs）中以1/1000孵育1小时，洗涤并转移至二氨基联*（.04%）和氯化镍（2.5%）中的0.1M乙酸钠和0.06%H2O2中6分钟。然后清洗、安装、干燥、脱水并盖上盖子。

根据我们的经验，如果按照上述方式储存和制备，如果您使用Vector精英试剂盒，每小瓶抗体应处理300至1000个30μm的白化大鼠脑（或类似大小的动物）切片。然而，你应该尝试集中注意力和程序，以确定哪一个最-适合你的环境。

Fluorochorome荧光金(Fluoro-Gold)抗体的检查和摄影：

使用宽带紫外（UV）激发滤光器，荧光显微镜观察荧光金。使用与在宽带紫外（如真蓝、快蓝、核黄）下激发的其他荧光逆行示踪剂相同的滤镜组，并应特别为荧光显微镜甘油或水制作物镜。由于塑料确实吸收紫外线，因此不建议通过塑料培养皿等进行观察。推荐使用T-Max（柯达，黑白）和Ektachrome 200（柯达，彩色幻灯片）。曝光时间通常从20秒到1.5分钟不等。

荧光金(Fluoro-Gold)抗体根据不同规格分为单管和二管：

荧光金抗体-单管/100ul（FC20001）

荧光金抗体-2管/2x100ul（FC20002）