Recombinant Human CUL4A/RBX1/DDB1/CRBN 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.
E3-650
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.
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Reconstitution Calculator
Background: CUL4A/RBX1/DDB1/CRBN Complex
Cullin-4A (CUL4A) is a core component of multiple cullin-RING type Ubiquitin ligase complexes that mediate the ubiquitination of proteins involved in cell cycle progression, DNA repair and other processes. In the DCX complex (DDB1-CUL4-X-box), CUL4A serves as a scaffold that organizes the DDB1-X-box substrate recognition subunits with the RBX1 subunit and contributes to catalysis through positioning of the substrate and an E2 ubiquitin-conjugating enzyme. In vivo, the E3 ubiquitin ligase activity of the DCX complex is dependent on neddylation of the CUL4 subunit, though neddylation may be dispensable in some in vitro reactions. This complex consists of an N-terminal 10-His tagged Cullin-4A, untagged RBX1, untagged DDB1 and untagged CRBN.
- Fischer E.S., et al. (2014) Nature 512: 49
- He Y.J., et al. (2006) Genes Dev. 20: 2949
- Ito T., et al. (2010) Science 327: 1345
- Wang H., et al. (2006) Mol. Cell 22: 383
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试题分析:酶是一种生物催化剂,在催化反应中本身不发生变化;此非酶的特性;B项体现了酶的作用条件较温和的特性;C项体现了酶的专一性;D项体现了酶的高效性;故选A。
考点:本题考查的是酶的特性。
点评:对于此类试题,学生应掌握酶的特性。
BAS-ELISA是在常规ELISA原理的基础上,结合生物素(B)与亲和素(A)间的高度放大作用,而建立的一种检测系统。生物素很易与蛋白质(如抗体等)以共价键结合。这样,结合了酶的亲和素分子与结合有特异性抗体的生物素分子产生反应,既起到了多级放大作用,又由于酶在遇到相应底物时的催化作用而呈色,达到检测未知抗原(或抗体)分子的目的。
辣根过氧化物酶(Horseradish Peroxidase,HRP)比活性高,稳定,分子量小,纯酶容易制备,所以最常用。HRP广泛分布于植物界,辣根中含量高,它是由无色的酶蛋白和棕色的铁卟啉结合而成的糖蛋白,糖含量18%。HRP由多个同功酶组成,分子量为40,000,等电点为PH3~9,酶催化的最适PH因供氢体不同而稍有差异,但多在PH5左右。酶溶于水和58%以下饱和度硫酸铵溶液。HRP的辅基和酶蛋白最大吸收光谱分别为403nm和275nm,一般以OD403nm /OD275nm的比值RZ(德文Reinheit Zahl)表示酶的纯度。高纯度的酶RZ值应在3.0左右(最高可达3.4)。RZ值越小,非酶蛋白就越多。 1) RZ>3 活性 >250u/mg, 主要应用于免疫学,是高纯度的过氧化酶。采用特殊色谱纯化技术以除去会影响免疫学反应的同工酶B .
2) RZ>2 活性 >180u/mg ,主要应用于临床化学,我们的客户也有将这个规格的产品应用于免疫学研究的。此时,一个标准化的分析方法就变得尤为重要。
3) RZ>1 活性 >100u/mg,主要应用于血糖试纸和尿液分析试纸。
4) RZ>0.6 活性 >60u/mg ,主要应用于尿液分析试纸。向左转|向右转
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