Using a straightforward RNA-isolation protocol, the CellAmp RT-PCR for Direct Prep Kit (Real Time) & Protein Analysis allows direct RNA template purification from animal cells for use in one- and two-step qRT-PCR. The kit enables RNA isolation from animal cells via a simple extraction method using animal cells cultured in 96-well (or other size well) plates. Direct RNA template purification from animal cells for use in real-time PCR takes approximately 10 minutes.
Using a straightforward RNA-isolation protocol, the CellAmp RT-PCR for Direct Prep Kit (Real Time) & Protein Analysis allows direct RNA template purification from animal cells for use in one- and two-step qRT-PCR. The kit enables RNA isolation from animal cells via a simple extraction method using animal cells cultured in 96-well (or other size well) plates. Direct RNA template purification from animal cells for use in real-time PCR takes approximately 10 minutes.
When used with a one-step real-time RT-PCR kit, theCellAmp Direct RNA Prep Kit (Real Time) & Protein Analysis allows gene expression analysis to be completed in approximately two hours. When combined with a reverse transcription reagent such as the PrimeScript RT Reagent Kit (Perfect Real Time), cDNA templates for real-time PCR can be synthesized in approximately 30minutes, enabling efficient high-throughput analyses. Direct RNA template purification can be accomplished using small batches of animal cells, a useful feature for gene expression profiling of precious or rare samples. The kit is also useful for studies of genes that are expressed at very low levels. Genomic DNA can be easily and efficiently removed using this kit. Finally, RNA template samples prepared withCellAmp Direct RNA Prep Kit (Real Time) & Protein Analysis are compatible with intercalating green dye and 5" nuclease probe-based assays for real-time PCR (qPCR), and thus can serve a wide variety of experimental applications.
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请问一般的实验室可以进行ELIspot吗？（我可以买到现成的试剂盒，资金这个应该没有问题。）ELIspot的难点在哪里？多谢大虾们的指点！这个对我很重要，多谢！！！
尤其是获得SCI文献引用的产品，
绝对ok的
用Mabteck公司的ALP标记抗体吧，配合Millipore公司的Multiscreen-IP96孔板，所有的试剂都可以减半，照样可以出非常好的结果，4*96孔的试剂盒我一直做8*96孔，但BCIP/NBT显色液没试过减半，因为它很便宜。记得当年第一次做实验就出了很好的结果，经过不断摸索，做出的蓝紫色斑点确实非常漂亮，几乎没有非特异斑点，嗯，ELISPOT是我最拿手的实验之一。
关于PBMC:
拿到血液标本后，首先我要做的是尽快分离PBMC（一般2h内），国产的分离液里天津灏洋的比较好，然后用冻存液冻存PBMC。PBMC的冻存过程比一般的细胞株要更为严格，最好用冻存器。ELISPOT试验的第一天，在包被抗体的同时，将细胞复苏，完全培养液培养过夜，第二天收集细胞并计数即可。
我想用酶联免疫斑点（ELISPOT）技术测定NK细胞活性，但不知道哪里能买到试剂盒，特向大家求助，谢谢。
