
easYmer HLA-A*24:02 MHC Tetramers Kit
easYmer HLA-A*24:02 MHC Tetramers Developed and Manufactured by immunAware
Materials Included:
The easYmer kit contains a peptide receptive preparation of HLA, a folding buffer, a positive control peptide for HLA-complex folding. The exact identities of these components are given below:
- easYmer: HLA-A*24:02 – peptide receptive and biotinylated HLA-A*24:02 molecules in Tris/Maleate pH7 with 30% Glycerol
- Folding Buffer: Tris/Maleate pH7
- Peptide: YF NS5 286-295 (KSEYMTSWFY), a known HLA-A*24:02 binder. Positive control for evaluation analysis of peptide-HLA folding
Available Sizes:
- 20 Tests (Sample-size)
- 50 Tests (Standard Size)
- 150 Tests
- 500 Tests
for Research Use Only
Key Benefits of easYmer® MHC Tetramers
- Ready-to-use
- One step loading
- Completely flexible and customizable*
- Biotinylated
- No special equipment needed
- Long shelf-life
*For Custom Tetramer Production, please Contact Us for more information
Assay Principle
This easYmer HLA-A*24:02 MHC Tetramers protocol is designed to evaluate the efficiency of peptide-HLA-I interaction and complex formation. The assay is based on detecting the ß2-microglobulin (ß2m? light chain subunit of recombinant HLA Class I (HLA-I) comlexes, where the heavy chain has been biotin tagged. These tagged complexes are subsequently captured by streptavidin coated beads, labeled with PE-conjugated anti-human ß2m, and analyzed by flow cytometry. Since peptide-HLA-I complex formation is entirely peptide dependent, bead-associated signals will only be detected if the peptide in question supports the folding of the HLA-I allotype of interest; peptides that efficiently support folding will give strong signals whereas peptides that support folding sub-optimally, or not at all, will give moderate to non-detectable signals.
Products Related to easYmer HLA-A*24:02 MHC Tetramers
easYmer HLA-A*24:07 MHC Tetramers Kit
easYmer HLA-A*25:01 MHC Tetramers Kit
easYmer HLA-A*30:02 MHC Tetramers Kit
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大家miRNA逆转录和qPCR的试剂盒用的是那个公司的啊,求推荐!
加尾法是采用加A酶先对mirna进行加尾,然后再用带oligodt的引物反转录,他的反转录引物是通用的,一次反转录可以获得所有miRNA的cdna,效率高。茎环法是采用特异性反转录引物序列+颈环结构作为反转录引物进行反转录的,一次反转录只能获得一种mirna的cdna,可能有几种一起反转录的,这个我不太清楚一起反转录的效果。然后茎环法的经典即ABI的探针法定量试剂盒。我用过茎环法反转录+染料法定量的,下游引物为通用引物,结果不太好,就现在用的这家的他采用的是双向引物都是特异性的,效果还可以。不知道我说的清楚不,希望能对你有帮助。
1.模板提取(一般为RNA):Trizol、氯仿、异丙醇、无水乙醇、DEPC处理水
2.模板浓度测定:分光光度计或NanoDrop
3.逆转录:逆转录试剂盒(或者一步法试剂盒),这一步可以用普通PCR做,也可以用水域做。
4.荧光定量PCR试剂:通常有用SYBR Green Mix做的,但是这里建议你用EvaGreen做,灵敏度和平行性都要好于SYBR Green,并且如果你那是ABI或者Stratagene的PCR如果用SYBR Green还需要加一步Rox很麻烦。
5.其他:除了以上的那些还需要离心管、PCR管或板(Axygen反应比较好)、移液枪等,暂时就想到这么多。
转录的只是插入的片断,怎么会跟模板一样大呢,一般情况下应该比模板小多了。我都是跑电泳,严格来说应该跑变性的;嫌麻烦可以非变性,快速(防止RNA降解)十分钟即可;抛出来的带会有两条,上面大的是模板,下面一条比较亮的是RNA,那么你的转录就 成功了。
PCR第一个步骤如果没记错的话应该叫“变性”, PCR的话是不需要解旋酶的,直接利用DNA的高温变性特性,在高温下,使氢键断开,DNA双链恢复为单链。 (还有解旋酶好像是解除双螺旋的结构,是一种拓扑异构酶,对于氢键的打开没有贡献吧
2.模板浓度测定:分光光度计或NanoDrop
3.逆转录:逆转录试剂盒(或者一步法试剂盒),这一步可以用普通PCR做,也可以用水域做。
4.荧光定量PCR试剂:通常有用SYBRGreenMix做的,但是这里建议你用EvaGreen做,灵敏度和平行性都要好于SYBRGreen,并且如果你那是ABI或者Stratagene的PCR如果用SYBRGreen还需要加一步Rox很麻烦。
5.其他:除了以上的那些还需要离心管、PCR管或板(Axygen反应比较好)、移液枪等,暂时就想到这么多。

