Recombinant Human UbcH7/UBE2L3 - Ubiquitin Charged Protein Summary
Pre-charged UBE2L3 may be added directly to in vitro reactions containing E3 Ubiquitin ligases and ligase substrates (if applicable)—no ATP, E1 enzyme, or extra Ubiquitin are required. Reaction conditions will need to be optimized for each specific application. Note: Reducing agents including dithiothreitol (DTT) or mercaptoethanol ( beta ME) may cause unintended thiolytic release of Ubiquitin from the complex—care must be taken if these compounds are present in buffers. We suggest using a thioester friendly reductant such as TCEP if possible.
26 kDa (18 kDa UBE2L3, 8.6 kDa Ubiquitin)
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.
E2-803
| Formulation | Approximately 0.45mg/ml (25 μM) with respect to UBE2L3. Formulated in 50 mM HEPES pH 7.5, 50 mM NaCl, 1 mM TCEP |
| 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.
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Reconstitution Calculator
Background: UbcH7/UBE2L3
Ubiquitin-conjugating Enzyme H7 (UbcH7), also known as Ubiquitin-conjugating Enzyme E2L 3 (UBE2L3), is a member of the Ubiquitinconjugating (E2) enzyme family. It has a predicted molecular weight of approximately 18 kDa. The human UbcH7 protein shares 100% amino acid (aa) sequence identity with the mouse and rat orthologs. UbcH7 has an E2 catalytic core domain that contains an active site cysteine residue and comprises 152 of its 154 aa residues. UbcH7 is catalytically active with HECT and RBR domaincontaining families of Ubiquitin ligases (E3s). UbcH7 localizes to both the nucleus and cytoplasm in human cells. In mice, its ortholog is expressed in many tissues including brain, muscle, heart, lung, lymph node, spleen, thymus, and testis. UbcH7 depletion results in an extended S phase and a reduced rate of proliferation, suggesting that it may play a role in the cell cycle. In humans, single nucleotide polymorphisms in UbcH7 are associated with systemic lupus erythematosus and Crohn"s disease, suggesting that UbcH7 is important for proper immune system function.
This product is an enzymatically generated UbcH7/UBE2L3-Ubiquitin thioester complex that has been purified to remove E1 Ubiquitin Activating enzyme, uncharged UbcH7/UBE2L3, free Ubiquitin, and Mg2+-ATP. The product provides a convenient starting material for use in single-turnover "Ubiquitin Discharge Assays," eliminating the need to either inhibit the E1 Ubiquitin Activating enzyme with potentially confounding chemical treatments or remove ATP via enzyme additions.
- Das, R. et al. (2013) EMBO J. 32: 2504
- Koyano, F. et al. (2014) Nature 510: 162
- Liew, C.W. et al. (2010) Biochem. J. 431: 23
- Petroski, M.D. & Deshaies, R.J. (2005) Cell 123: 1107
- Saha, A. et al. (2011) Mol. Cell 42: 75
FAQs
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为了使供输血用的血液能保存一定时间，加入适当的抗凝剂，并尽量防止在保存期发生变质而用的液体。为了防止因糖的消失引起活力降低，尚加入葡萄糖。ACD液（含柠檬酸三钠、柠檬酸、葡萄糖）一般用于血库。添加ACD的血液放在4—6℃条件下保存，可保存三周。另外为防止柠檬酸盐的过量可用加磷酸钠的CPD液。此外还设计了加乳酸钠的ACDL液。如果加入甘油等防冻剂则可长时间冷冻保存红细胞。
我搜索了一下网络上的，只找到说要保存在酸性下，但是没有具体的配方。
目前选择这个都是北京博凌科为生物科技有限公司的
小白一枚，现在要构建紫杉醇细胞耐药株。但是我们实验室紫杉醇是医用注射液，浓度为6mg/ml。这个浓度太大了，工作液浓度都是几十ng/ml。想问一下各位大神该如何配置成浓度相对低一点的储存液呢，有效期可以有多久呢？
目前正在从新鲜病人肿瘤组织标本中提取细胞悬液进行阳性细胞的筛选，但由于实验室距离医院有3个小时的车程，请问什么储存方法或者储存液能短期保证肿瘤组织中细胞完整性？谢谢
