![Signalchem/c-KIT (V559D), Active/5 ug/C06-12PG-05](images/Signalchem/smallvial.jpg)
Overview:
c-KIT is a proto-oncogene and a type 3 transmembrane receptor for MGF (mast cell growth factor, also known as stem cell factor). c-KIT was first identified as the cellular homolog of the feline sarcoma viral oncogene v-kit. c-KIT together with its ligand regulates growth and activation of a variety of hemopoietic and non-hemopoietic cells. Mutations in c-KIT are associated with gastrointestinal stromal tumors, mast cell disease, acute myelogenous lukemia, and piebaldism. Recently, deregulation of the KIT receptor TK by the prevalent activation loop mutation D816V has served as a focal point in therapeutic strategies aimed at curbing neoplastic mast cell growth (2).
Gene Aliases:
PBT, SCFR, CD117
Genbank Number:
NM_000222
References:
1. Berger, S A.: Signaling pathways influencing SLF and c-kit-mediated survival and proliferation. Immunol Res. 2006;35(1-2):1-12.2. Gotlib, J.: KIT mutations in mastocytosis and their potential as therapeutic targets. Immunol Allergy Clin North Am. 2006 Aug;26(3):575-92.
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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!!!
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。
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