MERS-CoV (NCoV / Novel coronavirus) Spike Protein fragment (RBD, aa 367-606) Insect Cell Lysate (WB positive control)

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MERS-CoV (NCoV / Novel coronavirus) Spike Protein fragment (RBD, aa 367-606) Insect Cell Lysate (WB positive control): 产品信息

产品描述
This MERS-CoV Coronavirus spike overexpression lysate was created in Baculovirus-Insect Cells and intented for use as a Western blot (WB) positive control. Purification of Coronavirus spike protein (Cat: 40071-V08B1) from the overexpression lysate was verified.
表达宿主
Baculovirus-Insect Cells
种属
MERS-CoV
蛋白构建信息
A DNA sequence encoding the MERS-CoV receptor binding domain (RBD) of spike protein fragment (AFS88936.1) (Glu367-Tyr606) was expressed with a polyhistidine tag at the C-terminus.
分子量
The recombinant receptor binding domain (RBD) of spike protein fragment MERS-CoV consists of 251 amino acids and predicts a molecular mass of 27.7 kDa. As a result of glycosylation, it migrates as an approximately 33.7 kDa band in SDS-PAGE under reducing conditions.

MERS-CoV (NCoV / Novel coronavirus) Spike Protein fragment (RBD, aa 367-606) Insect Cell Lysate (WB positive control): 使用指南

制备方法
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
裂解缓冲液
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
使用建议
1.  Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube. 2.  Re-dissolve the pellet using 200μL pure water and boil for 2-5 min.
缓冲液
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
稳定性 & 储存条件
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
应用
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

MERS-CoV (NCoV / Novel coronavirus) Spike Protein fragment (RBD, aa 367-606) Insect Cell Lysate (WB positive control): 别称

MERS-CoV coronavirus s1 Overexpression Lysate; MERS-CoV coronavirus s2 Overexpression Lysate; MERS-CoV coronavirus spike Overexpression Lysate; MERS-CoV cov spike Overexpression Lysate; MERS-CoV ncov RBD Overexpression Lysate; MERS-CoV ncov s1 Overexpression Lysate; MERS-CoV ncov s2 Overexpression Lysate; MERS-CoV ncov spike Overexpression Lysate; MERS-CoV RBD Overexpression Lysate; MERS-CoV S Overexpression Lysate; MERS-CoV s1 Overexpression Lysate; MERS-CoV Spike RBD Overexpression Lysate

Coronavirus spike 背景信息

The spike (S) glycoprotein of coronaviruses contains protrusions that will only bind to certain receptors on the host cell. Known receptors bind S1 are ACE2, angiotensin-converting enzyme 2; DPP4, dipeptidyl peptidase-4; APN, aminopeptidase N; CEACAM, carcinoembryonic antigen-related cell adhesion molecule 1; Sia, sialic acid; O-ac Sia, O-acetylated sialic acid. The spike is essential for both host specificity and viral infectivity. The term 'peplomer' is typically used to refer to a grouping of heterologous proteins on the virus surface that function together. The spike (S) glycoprotein of coronaviruses is known to be essential in the binding of the virus to the host cell at the advent of the infection process. It's been reported that SARS-CoV-2 (COVID-19 coronavirus, 2019-nCoV) can infect the human respiratory epithelial cells through interaction with the human ACE2 receptor. The spike protein is a large type I transmembrane protein containing two subunits, S1 and S2. S1 mainly contains a receptor binding domain (RBD), which is responsible for recognizing the cell surface receptor. S2 contains basic elements needed for the membrane fusion. The S protein plays key parts in the induction of neutralizing-antibody and T-cell responses, as well as protective immunity. The main functions for the Spike protein are summarized as: Mediate receptor binding and membrane fusion; Defines the range of the hosts and specificity of the virus; Main component to bind with the neutralizing antibody; Key target for vaccine design; Can be transmitted between different hosts through gene recombination or mutation of the receptor binding domain (RBD), leading to a higher mortality rate.
参考文献
  • Shen S, et al. (2007) Expression, glycosylation, and modification of the spike (S) glycoprotein of SARS CoV. Methods Mol Biol. 379: 127-35.
  • Du L, et al. (2009) The spike protein of SARS-CoV--a target for vaccine and therapeutic development. Nat Rev Microbiol. 7 (3): 226-36.
  • Xiao X, et al. (2004) The SARS-CoV S glycoprotein. Cell Mol Life Sci. 61 (19-20): 2428-30.

SARS-CoV-2 过表达细胞裂解液

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