Recombinant Human SUMO2 Fluorescein 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.
UL-756
| Formulation | Supplied as a solution in HEPES, DTT. |
| Shipping | The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage: | Protect from light. Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
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Reconstitution Calculator
Background: SUMO2
Human Small Ubiquitin-like Modifier 2 (SUMO2), also known as Sentrin2 and SMT3B is synthesized as a 95 amino acid (aa), propeptide with a predicted 11 kDa. SUMO2 contains a two aa C-terminal prosegment and an 18 aa N-terminal protein interacting region between aa 33-50. Human SUMO2 shares 100% aa sequence identity with mouse SUMO2. SUMO2 also has very high aa sequence identity with SUMO3 and SUMO4, 86% and 85%, respectively. SUMO2 shares only 44% aa sequence identity with SUMO1. SUMOs are a family of small, related proteins that can be enzymatically attached to a target protein by a post-translational modification process termed SUMOylation (1-3). All SUMO proteins share a conserved Ubiquitin domain and a C-terminal diglycine cleavage/attachment site. Following prosegment cleavage, the C-terminal glycine residue of SUMO2 is enzymatically attached to a lysine residue on a target protein. In humans, SUMO2 is conjugated to a variety of molecules in the presence of the SAE1/UBA2 SUMO-activating (E1) enzyme and the UBE2I/Ubc9 SUMO-conjugating (E2) enzyme (4,5). In yeast, the SUMO-activating (E1) enzyme is Aos1/Uba2p (6). Because of the high level of aa sequence identity most studies report effects of SUMO2/3. For example, post-translational addition of SUMO2/3 was shown to modulate the function of ARHGAP21, a RhoGAP protein known to be involved in cell migration (7). Other reports indicate that the SUMOylation with SUMO2/3, but not SUMO1, may represent an important mechanism to protect neurons during episodes of cerebral ischemia (8,9). However, studies suggest that SUMO2/3 expression is regulated in an isoform-specific manner since oxidative stress downregulated the transcription of SUMO3 but not SUMO2 (10).
SUMO-2 modified with fluorescein via primary amine coupling results in modification of lysine residues as well as the N-terminus. Although having a fully functional C-terminus, lysine modification may limit the ability of this reagent to propagate poly-SUMO chains. This labeled SUMO-2 allows for direct detection spectophotometrically with higher efficiency and sensitivity than with antibodies.
- Desterro, J.M. et al. (1997) FEBS. Lett. 417:297.
- Bettermann, K. et al. (2012) Cancer Lett. 316:113.
- Praefcke, G.J. et al. (2012) Trends Biochem. Sci. 37:23.
- Okuma, T. et al. (1999) Biochem. Biophys. Res. Commun. 254:693.
- Tatham, M.H. et al. (2001) J. Biol. Chem. 276:35368.
- Johnson, E.S. et al. (1997) EMBO J. 16:5509.
- Bigarella, C.L. et al. (2012) FEBS Lett. 586:3522.
- Datwyler, A.L. et al. (2012) J. Cereb. Blood Flow Metab. 31:2152.
- Wang, Z. et al. (2012) Protein Expr. Purif. 82:174.
- Sang, J. et al. (2012) Biochem. J. 435:489.
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谢谢各位！
不过制冰机里的冰不可以，那冰怎么得到呢？
protocol上面说在离心菌液的时候离心管里面要有1/2的冰啊，这个冰应该是碎冰而不是直接冻出来的整块的冰吧。
大家是怎么得到的呢？谢谢！！
制冰机原理：
制冰机是一种将水通过蒸发器由制冷系统制冷剂冷却后生成冰的制冷机械设备。根据蒸发器和生成过程方式原理不同，生成的冰的形状也不同，人们一般根据冰形状将制冰机分为颗粒冰机、片冰机、板冰机、管冰机、壳冰机等等。
制冰机的分类：
按冰的形状可分：鳞形片冰机，雪花制冰机，条冰机，板冰机，块冰机（可分食用小块冰机和工业用大块冰机），冰粒机，颗粒机，管冰机，子弹头制冰机。
实验室制冰机种类选择：
我们平时在外面就餐时，经常看到商家在饮料中加入冰块，有时候是方块的，有时候是中空的，还有在菜市场买海鲜的时候也会看到下面铺着一层冰，究竟有什么区别吗？我们在开展实验室工作的时候，应如何选择制冰机呢？
实验室制冰机主要是用于在开展生命科学相关实验时，给样品提供冰浴环境，防止样品（如核酸、蛋白）因为温度的变化导致降解或变质，从而保障实验结果的准确性和有效性。绝大部分的时候，我们的样品是放在EP管、离心管、pcr管等容器里面，然后把样品管插在冰浴中，靠接触来实现温度的保持。由于样品和冰无法直接接触，而是要通过管壁来传输，所以管壁和冰浴的接触面积是冰浴效果的保障，为了更好的保障样品，应尽量的扩大管壁和冰浴的接触面积。
针对以上考虑，实验室制冰机一般选择雪花制冰机，该种类型的制冰机生产出来的冰呈雪花状碎冰，由于颗粒很小，可以和管壁进行密切的接触，保障冰浴效果。
对于出冰的具体要求中，还有一点是非常重要的，出来的雪花冰要求有15-25%的含水量，这样可极大的提高冰浴和管壁的接触效果。
文章来源：厦门国仪科学仪器有限公司
