- SynonymINSR,Insulin receptor,IR,CD220
- SourceHuman Insulin R (28-944), His Tag (SPR verified) (INR-H52Ha) is expressed from human 293 cells (HEK293). It contains AA His 28 - Lys 944 (Accession # P06213-2).Predicted N-terminus: His 28Request for sequence
- Molecular Characterization
This protein carries a polyhistidine tag at the C-terminus.
The protein has a calculated MW of 106.5 kDa. The protein migrates as 115-125 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.
- EndotoxinLess than 1.0 EU per μg by the LAL method.
- Purity
>90% as determined by SDS-PAGE.
- 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.
Human Insulin R (28-944), His Tag (SPR verified) on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 90%.
Immobilized Human Insulin R (28-944), His Tag (Cat. No. INR-H52Ha) at 5 μg/mL (100 μL/well)can bind Human IGF-II, Fc Tag (Cat. No. IG2-H4260) with a linear range of 0.078-1.25 μg/mL (QC tested).
Immobilized Human Insulin R (28-944), His Tag (Cat. No. INR-H52Ha) at 5 μg/mL (100 μL/well)can bind Insulin Biotin Conjugate with a linear range of 0.02-0.313 μg/mL (Routinely tested).
Human Insulin R (28-944), His Tag (Cat. No. INR-H52Ha) captured on CM5 chip via anti-His antibody, can bind Human Insulin with an affinity constant of 249 nM as determined in a SPR assay (Biacore T200) (QC tested).
- BackgroundInsulin receptor (INSR) is also known as CD antigen CD220, which can be cleaved into the following 2 chains: Insulin receptor subunit alpha and Insulin receptor subunit beta. INSR is a tetramer of 2 alpha and 2 beta chains linked by disulfide bonds. The alpha chains carry the insulin-binding regions, while the beta chains carry the kinase domain. Forms a hybrid receptor with IGF1R, the hybrid is a tetramer consisting of 1 alpha chain and 1 beta chain of INSR and 1 alpha chain and 1 beta chain of IGF1R. In addition to binding insulin, the insulin receptor can bind insulin-like growth factors (IGFI and IGFII). Isoform Short of INSR has a higher affinity for IGFII binding. When present in a hybrid receptor with IGF1R, INSR binds IGF1.
- References
- (1)Pandini G., et al., 2002, J. Biol. Chem. 277:39684-39695.
- (2)Slaaby R., et al., 2006, J. Biol. Chem. 281:25869-25874.
- (3)Menting J.G., et al., 2013, Nature 493:241-245.
- (4)Kadowaki T., et al., 1988, Science 240:787-790.
Please contact us via TechSupport@acrobiosystems.com if you have any question on this product.
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2、可逆抑制剂：包括
a、竞争性抑制剂，抑制剂与底物竞争性结合酶反应中心，使Km增大，而Vmax不变 b、非竞争性抑制剂，酶与抑制剂结合后还能与底物结合，但活性降低，使Vmax减小，而Km不变
c、反竞争性抑制剂，酶只能与底物结合后才能与抑制剂结合，Vmax与Km都减小
可逆抑制剂可用透析等方法除去，使酶恢复作用
1、测定酶比活力：底物需要过量么？测定时间多长？是否可以加入过量的底物，然后测定3min吸光度的增加值，从吸光度的变化值计算比活力。
2、在酶抑制剂筛选的过程中，是否需要保证底物过量？还是要水浴一定时间让反应完全？我看到文献说用终浓度为0.1μmol/mL的底物，终浓度为45U/ml的酶，我想问，假设总体积为1ml，那么终浓度为45U/ml的酶岂不是每分钟能转化4.8μmol的底物？那么0.1μmol/mL的底物不就几秒钟就反应完了？那么怎么测定初速度？
《血管紧张素转换酶抑制剂在心血管病中应用的中国专家共识》.PDF(242.69k)
1.对于抑制剂筛选工作（求ic50）是不是体系内酶与底物的量（底物应该是过量的）对实验结果影响不大。
2.如果要求算Km值，是不是需要知道反应产物的绝对量。反应时间文献上都是5分钟，反应速度就用反应产物量除以反应时间即可。
3.酶是进口分装的，规格5U，一次用不完，用PBS稀释后如何保存
谢谢
