Recombinant Human CUL2/NEDD8/RBX1 Complex Protein, CF Summary
Product Datasheets
Carrier Free
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins.Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration.The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard.In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
E3-421
Formulation | X mg/ml (XμM) in 50 mM HEPES pH 7.5, 200 mM NaCl, 10% (v/v) Glycerol, 1 mM DTT |
Shipping | The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below. |
Stability & Storage: | Use a manual defrost freezer and avoid repeated freeze-thaw cycles. |
Reconstitution Calculator
Background: CUL2/NEDD8/RBX1 Complex
Cullin-2 is a core component of multiple cullin-RING based "ECS" (Elongin B/C-CUL2-SOCS-box protein) E3 Ubiquitin ligase complexes that mediate the ubiquitination of proteins involved in a variety of cellular processes. In the ECS complex, Cullin-2 serves as a scaffold that organizes the ELOB/C-associated recognition subunits with the RBX1 subunit and contributes to the catalysis through positioning of the substrate and an E2 ubiquitin-conjugating enzyme. In vivo, the E3 ubiquitin ligase activity of the ECS complex is dependent on neddylation of the cullin subunit, though neddylation may be dispensable for some in vitro reactions. This complex consists of an N-terminal 10-His tagged Cullin-2 (UniProt Q13617) and untagged RBX1 (P62877). CUL2 is neddylated in this complex.
- Baek K., et al. (2020) Nature 578: 461
- Duda D.M., et al. (2008) Cell 134: 995
- Duda D.M., et al. (2012) Mol. Cell 47: 371
- Goldenberg S.J., et al. (2004) Cell 119: 517
- Zheng N., et al. (2002) Nature 416: 703
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只要是细胞,就得走向衰老,现代研究表明,细胞的衰老和端粒的变短有关系。端粒就是DNA,随着细胞分裂次数的增多,端粒在不断变短,端粒酶就是组织端粒变短的。然而正常细胞中,端粒酶的活性受抑制,端粒酶是怎么染端粒变长的呢?实际上端粒酶是由RNA组成的,可以根据碱基互补配对逆转形成DNA,使得变短的端粒变长。所以说端粒酶可以逆转录形成端粒,使端粒不减短!
各位大侠:小弟最近做个长片段的CDNA与T载体的连接,片段大小6.4K,载体选用的是pUC19和pGEMT-easy,反转录酶用的是RevertAidTirstStrandcDNASynthesisKit(#K1621),CDNA第二链的扩增用的是LATaq。我是用纯病毒的RNA做模板,操作步骤严格按照说明做的,也曾经有一次获得过电泳图清晰显示的是全长6.4KB左右的片段,与载体连接,结果筛选到的几个克隆,插入的片段仅有2KB左右,奈何?可最近几次再用此反转录试剂盒却不能得到所需片段。在此,小弟想问问大家,你们是否遇到同样的问题,是如何解决的?所用的试剂(载体和酶的选择)和方法是什么,可以告诉后来者,共同进步吧。
我在做5RACE,选用TaKaRa公司的RNA反转录酶(RNaseH-),不知道这个酶好不好,有什么需要注意的。
2. 禽成髓细胞瘤病毒(AMV)反转录酶:有强的聚合酶活性和RNA酶H活性。最适作用温度为42℃。
3.Thermus thermophilus、Thermus flavus等嗜热微生物的热稳定性反转录酶:在Mn存在下,允许高温反转录RNA,以消除RNA模板的二级结构。
4.MMLV反转录酶的RNase H突变体:商品名为SuperScript 和SuperScriptⅡ。此种酶较其它酶能多将更大部分的RNA转换成cDNA,这一特性允许从含二级结构的、低温反转录很困难的mRNA模板合成较长cDNA。向左转|向右转