Product Description:

DKK-1 is a member of the DKK protein family which also includes DKK-2, DKK-3 and DKK-4. DKK-1 was originally identified as a Xenopus head-forming molecule that behaves as an antagonist for Wnt signaling. Subsequent studies have shown that DKK-1 and DKK-4 play an important regulatory role in the Wnt/β-catenin signaling pathway by forming inhibitory complexes with LDL receptor-related proteins 5 and 6 (LRP5 and LRP6), which are essential components of the Wnt/β-catenin signaling system. LPR5 and LPR6 are single-pass transmembrane proteins that appear to act as co-receptors for Wnt ligands involved in the Wnt/β-catenin signaling cascade. It has been suggested that by inhibiting Wnt/β-catenin signaling, which is essential for posterior patterning in vertebrates, DKK-1 permits anterior development. This notion is supported by the finding that mice deficient of DKK-1 expression lack head formation and die during embryogenesis. Mature human DKK-1 expressed in HEK293 cells is a 35-40 kDa glycoprotein containing 235 amino acid residues.

Technical Specifications:

• Species: Human
• Expression: HEK293 Cell Expressed
• Purity: ≥97%
• Endotoxin: <1.0 EU/μg
• Molecular Mass: 25.8 kDa
• Formulation: 1x PBS, pH 7.4
• Country of Origin: USA

Purity Confirmation:

This was determined by SDS-PAGE gel and HPLC analysis.

Activity Assay:

Determined by its ability to inhibit the proliferation of HCT116 colorectal carcinoma cells. Approximately 40% growth inhibition was achieved at a DKK-1 concentration of 200ng/ml.

AA Sequence:

TLNSVLNSN	IKNLPPPLGG	AAGHPGSAVS
AAPGILYPGG	NKYQTIDNYQ	PYPCAEDEEC
GTDEYCASPT	RGGDAGVQIC	LACRKRRKRC
MRHAMCCPGN	YCKNGICVSS	DQNHFRGEIE
ETITESFGND	HSTLDGYSRR	TTLSSKMYHT
KGQEGSVCLR	SSDCASGLCC	ARHFWSKICK
PVLKEGQVCT	KHRRKGSHGL	EIFQRCYCGE
GLSCRIQKDH	HQASNSSRLH	TCQRH

Reconstitution Buffer:

Centrifuge the vial prior to opening. Reconstitute in water to a concentration of 0.1-1.0 mg/ml. Do not vortex.

Storage:

For extended storage, it is recommended to further dilute in a buffer containing a carrier protein (example 0.1% HSA) and store in working aliquots at -20°C to -80°C.

HumanZyme始建于2005年。世界上首家从人细胞生产人类逼真蛋白的知名公司。产品包括：人源化的细胞因子、及相关的抗体、人源化的重组白蛋白、和树突细胞培养试剂盒。产品行销到世界各地，产品受到广泛的好评。并与世界著名经销商有长期的供应合同。HumanZyme利用自己的专利技术，突破了人细胞宿主表达人类蛋白低的瓶颈，研制成了既能贴壁生长又能悬浮培养的细胞系。这不仅可以供应实验室规模量，而且可以大规模GMP生产，以满足临床前和临床需求。HumanZyme 是独家可以提供无动物源污染和无异源蛋白污染的公司。2 蛋白表达的背景（以人细胞和大肠杆菌细胞为例）：蛋白质的合成是以DNA为模板通过转录、翻译和翻译后的修饰，蛋白才能有其生物结构和功能，最终到达靶器官才能起到生物学效应。特别是翻译后的修饰，如折叠、糖基化、磷酸化、等都是在细胞器中进行。如下表表所示，大肠杆菌内没有细胞器。翻译后的修饰多为人工修饰，其结构和自然结构有很大距离。人细胞和细菌的结构比较细胞器人细胞宿主大肠杆菌宿主高尔基复合体有无内质网有无线粒体有无中心体有无溶酶体有无EcoliHuman3与大肠杆菌和其他宿主表达的蛋白比，HumanZyme产品优势：i. 工程药物合成来源于无支原体和病毒污染，有着良好安全记录的人的HEK293，经过长时间的筛选和比较，既易于转染、更能大量表达目的蛋白的细胞HEK293TS。ii. 采用无血清(Serum-free)，无蛋白(Protein-free)，化学成分确定(Chemically Defined)的组合培养基，有效避免了血清中复杂的蛋白成分，如动物白蛋白(albumin)，动物转铁蛋白(transferrin)，动物胰岛素(Insulin)以及类胰岛素生长因子（IGF）对细胞和终产品的污染。iii. 创建特别适合人源细胞的高表达载体(Expression Vector)，由人源超强启动子(Human Promoter)和信号肽(Human Signal Peptide)驱动。相对于其它表达系统利用的病毒驱动子例如CMV和 SV40，从安全和有效性上有了更好的保障。iv. 重组蛋白质分子无标记(Tag-free)，专有的纯化手段步骤少，效率高。v.  无异源性（xeno-free）、无动物源性（animal-free）、超低毒素、纯度高、无其他蛋白载体（如BSA）。vi. 可以大量生产细菌宿主不能表达的系列蛋白，如转化生长因子超家族系列（TGFβ super families）。vii. 逼真的糖基化、磷酸化、和正确的折叠。从而体外的半衰期长、稳定性高。（如DC-KIT，一星期免换液，可与同类产品比美）4 产品的应用领域：a) 干细胞和再生医学领域的研究，如人胚胎干细胞（hESCs）、可诱导多能干细胞（iPS）、间充质干细胞（MSC）等的扩增与分化的理想产品。如Activin A、TGFs、BMPs、FGFs、Noggin等。b) 微生物和免疫研究：如白介素和炎症因子系列产品，如IL1、IL4、IL6、IL17s、IL23、TNFα、c) 细胞培养基的原料添加物，如碱性成纤维细胞生长因子（FGFbasic）和转铁蛋白（transferrin）等。d) 细胞生物治疗和生物医药治疗公司，干扰素、生血因子等。