Advanced negative detection method comparable to silver stain for SDS-PAGE separated proteins detection

2016 ◽  
Vol 510 ◽  
pp. 21-25 ◽  
Author(s):  
Xu Wang ◽  
Sun-Young Hwang ◽  
Wei-Tao Cong ◽  
Li-Tai Jin ◽  
Jung-Kap Choi
Keyword(s):  
Detection Method ◽  
Silver Stain ◽  
Sds Page ◽  
Negative Detection

scholarly journals High-throughput negative detection of SDS-PAGE separated proteins and its application for proteomics

2010 ◽  
Vol 31 (2) ◽  
pp. 411-420 ◽  
Author(s):  
Wei-Tao Cong ◽  
Sun-Young Hwang ◽  
Li-Tai Jin ◽  
Hong-Zhang He ◽  
Jung-Kap Choi
Keyword(s):  
High Throughput ◽  
Sds Page ◽  
Negative Detection

An expanding negative detection method for the visualization of DNA in polyacrylamide gels by eosin Y

2010 ◽  
Vol 31 (20) ◽  
pp. 3450-3456 ◽  
Author(s):  
Zhong-Xin Zhu ◽  
Wei-Tao Cong ◽  
Peng-Hui Zhang ◽  
Wei-De Ma ◽  
Liu Min ◽  
...  
Keyword(s):  
Detection Method ◽  
Eosin Y ◽  
Polyacrylamide Gels ◽  
Negative Detection

A novel electrochemical DNA detection method based on bio-barcode/gold label silver stain dual amplification

2020 ◽  
Vol 10 (2) ◽  
pp. 206-213
Author(s):  
Pengfei Jiang ◽  
Miduo Tan ◽  
Lan Zhao ◽  
Yulin Wang ◽  
Ting Pan ◽  
...  
Keyword(s):  
Electrochemical Detection ◽  
Detection Method ◽  
Linear Sweep Voltammetry ◽  
Silica Nanoparticle ◽  
Silver Ions ◽  
Silver Deposition ◽  
Silver Stain ◽  
Modified Dna ◽  
Gold Label ◽  
Dual Amplification

A novel electrochemical gene detection method based on bio-barcode/gold label silver stain (GLSS) dual amplification is presented. The silica nanoparticle/gold nanoparticles (SiO2/AuNPs) barcode probes were constructed by combining SiO2 nanoparticle-labeled DNA with AuNPs, which hybridizes with target DNAs and the electrode surface-modified DNA probes. The silver ions were reduced on the surface of AuNPs to form a silver shell by the catalysis of AuNPs, and then the electrochemical detection of deposited silver was performed by linear sweep voltammetry (LSV). In this paper, the effects of electrode blocking agent, blocking time and silver deposition time on electrochemical detection were also investigated. The proposed electrochemical DNA biosensor bring forth a low detection limit of 0.23 fM and a widely linear range of 1 fM to 10 pM. This method can clearly distinguish the complementary and mismatched DNA, with good repeatability and stability.

scholarly journals Erratum: High-throughput negative detection of SDS-PAGE separated proteins and its application for proteomics

2010 ◽  
Vol 31 (16) ◽  
pp. 2858-2858
Author(s):  
Wei-Tao Cong ◽  
Sun-Young Hwang ◽  
Li-Tai Jin ◽  
Hong-Zhang He ◽  
Jung-Kap Choi
Keyword(s):  
High Throughput ◽  
Sds Page ◽  
Negative Detection

scholarly journals Produksi imunoglobulin Y (IgY) untuk pengembangan metode deteksi dini kontaminasi okratoksin (Immunoglobulin Y (IgY) production to develop an early detection method for ochratoxin contamination)

2018 ◽  
Vol 86 (1) ◽  
Author(s):  
Irma KRESNAWATY ◽  
. SUHARYANTO ◽  
. SISWANTO ◽  
Sumi HUDIYONO
Keyword(s):  
Early Detection ◽  
Ochratoxin A ◽  
Immunosorbent Assay ◽  
Detection Method ◽  
Polyclonal Antibodies ◽  
Egg Laying ◽  
Enzyme Linked Immunosorbent Assay ◽  
Dot Blot ◽  
Negative Effects ◽  
Sds Page

Indonesian coffee and cocoa commodities are constrained by low product quality problem due to contamination of fungal metabolites which  producing ochratoxin A (OTA). Ochratoxin is neprotoxic, immunogenic, carcinogenic and teratogenic to the human health. Early detection method on site detection should be developed  because of  those negative effects. The aim of this study  was to produce antibody to develop a method for  OTA detection. Antibody was produced by immunization of egg laying hen. Antibody-produced was sepatared and analyzed using ELISA (Enzyme-Linked Immunosorbent Assay) and DBIA (dot blot immunoassay),and tested its composition using HPLC and SDS PAGE. The results showed that anti-OTA polyclonal antibodies had been obtained already from chicken eggs in the 4th period (7 weeks after initial immunization). These antibodies showed anti-OTA reactivity by DBIA method and still showed anti-OTA reactivity up to 9th period (12 weeks after initial immunization). The anti-BSA antibodies produced should be removed to increase the sensitivity of antibodies againts ochratoxin A. The separation of BSA antibodies can be conducted by the absorption of the protein.  [Keywords: ochratoxin A; early detection; antibody IgY]. AbstrakKomoditas kopi dan kakao Indonesia terkendala masalah mutu produk yang rendah akibat kontaminasi cendawan penghasil okratoksin A. Okratoksin A (OTA) bersifat neprotoksik, imunogenik, karsinogenik dan teratogenik yang membahayakan kesehatan manusia. Karena efek negatif yang diakibatkan oleh mikotoksin ini, maka perlu dikembangkan deteksi dini kontaminasi okratoksin langsung di lokasi. Penelitian ini bertujuan menghasilkan antibodi imunoglobulin Y (IgY) untuk mengembangkan metode perakitan perangkat deteksi cepat berbasis imunologi untuk deteksi OTA. Antibodi dihasilkan menggunakan uji ayam petelur. Antibodi yang dihasilkan dipisahkan dan dianalisis aktivitasnya dengan ELISA (Enzyme-Linked Immunosorbent Assay) dan DBIA (dot blot immunoassay), serta diuji komposisinya dengan HPLC dan SDS PAGE. Hasil penelitian menunjukkan bahwa antibodi poliklonal anti-OTA sudah diperoleh dari telur ayam pada periode ke-4 (7 minggu setelah imunisasi awal). Antibodi ini menunjukkan reaktivitas anti-OTA dengan metode DBIA dan masih menunjukkan reaktivitas anti-OTA sampai periode 9 (12 minggu setelah imunisasi awal). Komposisi asam amino antibodi anti-OTA menunjukkan perbedaan dengan komposisi asam amino IgY di database. Antibodi anti BSA yang dihasilkan harus dihilangkan terlebih dahulu untuk meningkatkan sensitivitas antibodi terhadap okratoksin A dan pemisahan dapat dilakukan dengan penyerapan antibodi BSA.[Kata Kunci:  okratoksin A;  deteksi dini; antibodi IgY].

3-D organization of fibrinogen receptors using computer reconstruction and whole-mount microscopy

1988 ◽  
Vol 46 ◽  
pp. 30-31
Author(s):  
E. T. O'Toole ◽  
R. R. Hantgan ◽  
J. C. Lewis
Keyword(s):  
Whole Blood ◽  
Colloidal Gold ◽  
Blood Platelets ◽  
Cell Contact ◽  
Computer Assisted ◽  
Adherent Cells ◽  
Differential Centrifugation ◽  
Computer Reconstruction ◽  
Sds Page ◽  
Whole Mount

Thrombocytes (TC), the avian equivalent of blood platelets, support hemostasis by aggregating at sites of injury. Studies in our lab suggested that fibrinogen (fib) is a requisite cofactor for TC aggregation but operates by an undefined mechanism. To study the interaction of fib with TC and to identify fib receptors on cells, fib was purified from pigeon plasma, conjugated to colloidal gold and used both to facilitate aggregation and as a receptor probe. Described is the application of computer assisted reconstruction and stereo whole mount microscopy to visualize the 3-D organization of fib receptors at sites of cell contact in TC aggregates and on adherent cells.Pigeon TC were obtained from citrated whole blood by differential centrifugation, washed with Ca++ free Hank's balanced salts containing 0.3% EDTA (pH 6.5) and resuspended in Ca++ free Hank's. Pigeon fib was isolated by precipitation with PEG-1000 and the purity assessed by SDS-PAGE. Fib was conjugated to 25nm colloidal gold by vortexing and the conjugates used as the ligand to identify fib receptors.

Solid-phase immunoelectron microscopy for rapid diagnosis of enteroviruses

1984 ◽  
Vol 42 ◽  
pp. 226-227 ◽  
Author(s):  
K. Pegg-Feige ◽  
F. W. Doane
Keyword(s):  
Electron Microscopy ◽  
Cell Culture ◽  
Immunoelectron Microscopy ◽  
Detection Method ◽  
Solid Phase ◽  
Protein A ◽  
Plant Viruses ◽  
Rapid Diagnosis ◽  
Virus Diagnosis ◽  
Antiviral Antibody

Immunoelectron microscopy (IEM) applied to rapid virus diagnosis offers a more sensitive detection method than direct electron microscopy (DEM), and can also be used to serotype viruses. One of several IEM techniques is that introduced by Derrick in 1972, in which antiviral antibody is attached to the support film of an EM specimen grid. Originally developed for plant viruses, it has recently been applied to several animal viruses, especially rotaviruses. We have investigated the use of this solid phase IEM technique (SPIEM) in detecting and identifying enteroviruses (in the form of crude cell culture isolates), and have compared it with a modified “SPIEM-SPA” method in which grids are coated with protein A from Staphylococcus aureus prior to exposure to antiserum.

Immunocytochemical detection of hepatic carbohydrate metabolic enzymes: Improved resolution with polyethylene glycol embedding and visio-bond semithin sections

1992 ◽  
Vol 50 (1) ◽  
pp. 792-793
Author(s):  
Kuixiong Gao ◽  
Randal E. Morris ◽  
Bruce F. Giffin ◽  
Robert R. Cardell
Keyword(s):  
Polyethylene Glycol ◽  
Glycogen Phosphorylase ◽  
Phosphoenolpyruvate Carboxykinase ◽  
Glycogen Synthase ◽  
Metabolic Enzymes ◽  
Silver Stain ◽  
Accurate Localization ◽  
Immunocytochemical Detection ◽  
Semithin Sections ◽  
Parenchymal Cells

Several enzymes are involved in the regulation of anabolic and catabolic pathways of carbohydrate metabolism in liver parenchymal cells. The lobular distribution of glycogen synthase (GS), phosphoenolpyruvate carboxykinase (PEPCK) and glycogen phosphorylase (GP) was studied by immunocytochemistry using cryosections of normal fed and fasted rat liver. Since sections of tissue embedded in polyethylene glycol (PEG) show good morphological preservation and increased detectability for immunocytochemical localization of antigenic sites, and semithin sections of Visio-Bond (VB) embedded tissue provide higher resolution of cellular structure, we applied these techniques and immunogold-silver stain (IGSS) for a more accurate localization of hepatic carbohydrate metabolic enzymes.

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