產品描述This 2019-nCoV CoV spike overexpression lysate was created in Baculovirus-Insect Cells and intented for use as a Western blot (WB) positive control. Purification of CoV spike protein from the overexpression lysate was verified.
表達宿主Baculovirus-Insect Cells
種屬2019-nCoV
蛋白構建信息A DNA sequence encoding the NCP-CoV(2019-nCoV) Spike Protein (S1 Subunit) was expressed with a polyhistidine tag at the C-terminus.
分子量The recombinant NCP-CoV(2019-nCoV) Spike Protein (S1 Subunit, His tag) consists of 681 amino acids and predicts a molecular mass of 76.45 kDa.
SARS-CoV-2 (2019-nCoV) Spike Insect Cell Lysate (WB positive control): 使用指南
制備方法Cell lysate was prepared by homogenization 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.
裂解緩沖液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.
使用建議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. 3. Store the lyophilized cell lysate at 4℃. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80℃.
緩沖液1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
穩定性 & 儲存條件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.
應用Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.
SARS-CoV-2 (2019-nCoV) Spike Insect Cell Lysate (WB positive control): 別稱
2019-nCoV coronavirus spike Overexpression Lysate; 2019-nCoV cov spike Overexpression Lysate; 2019-nCoV ncov RBD Overexpression Lysate; 2019-nCoV ncov s1 Overexpression Lysate; 2019-nCoV ncov s2 Overexpression Lysate; 2019-nCoV ncov spike Overexpression Lysate; 2019-nCoV NCP-CoV RBD Overexpression Lysate; 2019-nCoV NCP-CoV s1 Overexpression Lysate; 2019-nCoV NCP-CoV s2 Overexpression Lysate; 2019-nCoV NCP-CoV Spike Overexpression Lysate; 2019-nCoV novel coronavirus RBD Overexpression Lysate; 2019-nCoV novel coronavirus s1 Overexpression Lysate; 2019-nCoV novel coronavirus s2 Overexpression Lysate; 2019-nCoV novel coronavirus spike Overexpression Lysate; 2019-nCoV RBD Overexpression Lysate; 2019-nCoV S1 Overexpression Lysate; 2019-nCoV S2 Overexpression Lysate; 2019-nCoV Spike RBD Overexpression Lysate
CoV 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 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.
研究領域
Microbiology>>Pathogenic microorganism>>viruses>>Animal virus>>RNA virus>>+ssRNA virus>>Coronaviridae>>Coronavirus (CoV)>>CoV viral antigen
參考文獻
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.

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