
Product Description
The Gold-in-a-Box™ Conjugation Kit, 30 nm and 40 nm, contains the reagents required to prepare highly-reactive antibody or soluble protein gold conjugates that can be utilized in lateral-flow assays.
The key to most high-performance lateral-flow assays is the ability to attach antibodies/proteins to gold nanoparticles. The Gold-in-a-Box™ Conjugation Kit enables optimal binding of an antibody/protein to our Naked Gold 30 nm and 40 nm highly concentrated gold sols, while retaining a high degree of specific activity through adjusting the pH of the gold sols to slightly above the isoelectric point of the coating antibody/protein. This is done through a series of pH titrations with the provided buffers. The Gold-in-a-Box™ kit allows researchers and assay developers to quickly (in less than 50 minutes) determine the pI and optimal coating conditions for your antibody or soluble protein and to perform the conjugation reaction. A sample preparation and gold conjugation procedure is provided.
Reagents are also provided to coat the resultant gold conjugates on polyester of glass fiber ribbon.
Specifications:
Kit Contents:
- NG30-B009 – Naked Gold 30 nm Concentrated Gold Sol at 15 O.D./mL, 9 mL
- NG40-B009 – Naked Gold 40 nm Concentrated Gold Sol at 15 O.D./mL, 9 mL
- BUFA-001 – Buffer Solution A, 1.0 mL
- BUFB-001 – Buffer Solution B, 1.0 mL
- BUFC-001 – Buffer Solution C, 1.0 mL
- BUFD-001 – Buffer Solution D, 1.0 mL
- BLK-002 – BSA Blocking Stabilizer Solution, 2.0 mL
- CDB-002 – Gold Drying Buffer, 2.0 mL
Storage Temperature: Refrigerated (2-8°C), do not freeze
Shipping Temperature: Ambient temperatures
Stability: 2 years from date of manufacture
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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。

