- SynonymEGFRvIII
- SourceBiotinylated Human EGFRvIII, Avitag,His Tag (EGR-H82E0) is expressed from human 293 cells (HEK293). It contains AA Leu 25 - Ser 378 (Accession # NP_001333870.1).Predicted N-terminus: Leu 25Request for sequence
- Molecular Characterization
This protein carries an Avi tag (Avitag™) at the C-terminus, followed by a polyhistidine tag.
The protein has a calculated MW of 42.2 kDa. The protein migrates as 60-90 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.
- BiotinylationBiotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.
- Biotin:Protein RatioThe biotin to protein ratio is 0.5-1 as determined by the HABA assay.
- EndotoxinLess than 1.0 EU per μg by the LAL method.
- Purity
>95% as determined by SDS-PAGE.
>90% as determined by SEC-MALS.
- Formulation
Lyophilized from 0.22 μm filtered solution in PBS, pH7.4. Normally trehalose is added as protectant before lyophilization.
Contact us for customized product form or formulation.
- Reconstitution
Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
- Storage
For long term storage, the product should be stored at lyophilized state at -20°C or lower.
Please avoid repeated freeze-thaw cycles.
This product is stable after storage at:
- -20°C to -70°C for 12 months in lyophilized state;
- -70°C for 3 months under sterile conditions after reconstitution.
Biotinylated Human EGFRvIII, Avitag,His Tag on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 95%.
The purity of Biotinylated Human EGFRvIII, Avitag,His Tag (Cat. No. EGR-H82E0) was more than 90% and the molecular weight of this protein is around 60-70 kDa verified by SEC-MALS.
Immobilized Biotinylated Human EGFRvIII, Avitag,His Tag (Cat. No. EGR-H82E0) at 1 μg/mL (100 μL/well) on streptavidin (Cat. No. STN-N5116) precoated (0.5 μg/well) plate, can bind Anti-EGFRvIII Antibody , Human IgG1 with a linear range of 0.1-3 ng/mL (QC tested).
Immobilized CetuxiMab at 1 μg/mL (100 μL/well) can bind Biotinylated Human EGFRvIII, Avitag,His Tag (Cat. No. EGR-H82E0) with a linear range of 0.2-2 ng/mL (Routinely tested).
- Citations
Small-molecule probes for affinity-guided introduction of biocom-patible handles on metal-binding proteins
Authors: Mortensen MR, et al.
Journal: Bioconjug Chem 2018
Application: SPR
Request for Full-text
- Amino Acid Sequence
LEEKKGNYVVTDHGSCVRACGADSYEMEEDGVRKCKKCEGPCRKVCNGIGIGEFKDSLSINATNIKHFKNCTSISGDLHILPVAFRGDSFTHTPPLDPQELDILKTVKEITGFLLIQAWPENRTDLHAFENLEIIRGRTKQHGQFSLAVVSLNITSLGLRSLKEISDGDVIISGNKNLCYANTINWKKLFGTSGQKTKIISNRGENSCKATGQVCHALCSPEGCWGPEPRDCVSCRNVSRGRECVDKCNLLEGEPREFVENSECIQCHPECLPQAMNITCTGRGPDNCIQCAHYIDGPHCVKTCPAGVMGENNTLVWKYADAGHVCHLCHPNCTYGCTGPGLEGCPTNGPKIPSGLNDIFEAQKIEWHEHHHHHH
- BackgroundThe epidermal growth factor receptor (EGFR; ErbB-1; HER1 in humans) is the cell-surface receptor for members of the epidermal growth factor family (EGF-family) of extracellular protein ligands. The epidermal growth factor receptor is a member of the ErbB family of receptors, a subfamily of four closely related receptor tyrosine kinases: EGFR (ErbB-1), HER2/c-neu (ErbB-2), Her 3 (ErbB-3) and Her 4 (ErbB-4). Mutations affecting EGFR expression or activity could result in cancer.The type III EGF deletion-mutant receptor (EGFRvIII) is the most common mutation and was first identified in primary human glioblastoma tumors; EGFR gene amplification is correlated with the structural rearrangement of the gene. The EGFRvIII gene has an in-frame deletion of 801 base pairs, corresponding to exons 2–7 in the mRNA, resulting in the deletion of amino acids 30-297 in the extracellular domain and the generation of a glycine at the fusion point.
- References
Please contact us via TechSupport@acrobiosystems.com if you have any question on this product.
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酶标仪按照功能的不同划分,可以分为（1）光吸收酶标仪（可见酶标仪，紫外/可见酶标仪）（2）荧光酶标仪（3）化学发光酶标仪。
分光光度计按照波长及应用领域的不同可以分为：（1）可见光分光光度计（2）紫外分光光度计（3）红外分光光度计（4）荧光分光光度计（5）原子吸收分光光度计。
分光光度计和酶标板存在一些不同之处，具体差别体现在以下三个方面：
（1）盛装待测溶液的容器：
分光光度计用的是比色皿，酶标仪使用的是塑料微孔板(酶标板)。比色皿只能起到盛装溶液的作用，每个比色皿一次只能盛装一种溶液。酶标板常用透明的聚乙烯材料制成，对抗原抗体有较强的吸附作用，因此用它作为固相载体，酶标板通常为48孔或96孔，每个微孔可以盛装不同的溶液。
（2）光路的方向：
分光光度计是水平光路，而酶标仪则是垂直光路。由于酶标板盛样本的塑料微孔板是多排多孔的,光线只能垂直穿过,因此酶标仪的光束都是垂直通过待测溶液和微孔板的,光束既可是从上到下,也可以是从下到上穿过比色液。垂直光的特点是标本吸光度受液体浓缩或稀释的影响小，不足之处是受被测样本液面是否水平、酶标板透光性、孔底是否平整等的影响较大。
（3）光路的长度：
由于光密度（OD值）与吸光系数, 待测组分的浓度以及光路长度成正比关系。
分光光度计采用的比色杯的宽度通常是1cm，所以光路长度固定为1cm。因此不同仪器，不同批次测量的数据具有同样的可比性。
而酶标仪采用的是垂直光路, 所以光路的长度应该是液体液面的高度。所以测得的值受到样品的体积的影响。
请问BAC蛋白浓度测定波长是562，我们这边酶标仪的滤光片波长最接近的是595，这样也可以测吗
各位前辈，MTT法加样后第一天阳性组镜下看细胞全死了，样品组和阴性组镜下状态还可以，加DMSO后样品孔和阴性孔颜色粉红，阳性孔红紫到发黑，酶标仪检测，样品组和阴性组大概0.2-0.4，阳性组却有1.2-1.4，请问，这是怎么回事？细胞接板密度为4*104,100μL。向各位求助了，万分感谢！
