Development of eastern blotting technique for sennoside A and sennoside B using anti-sennoside A and anti-sennoside B monoclonal antibodies

2009 ◽  
Vol 20 (2) ◽  
pp. 154-158 ◽  
Author(s):  
Osamu Morinaga ◽  
Takuhiro Uto ◽  
Seiichi Sakamoto ◽  
Waraporn Putalun ◽  
Sorasak Lhieochaiphant ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Eastern Blotting ◽  
Blotting Technique

Monoclonal antibodies can precipitate low-density lipoprotein. I. Characterization and use in determining apolipoprotein B.

1985 ◽  
Vol 31 (10) ◽  
pp. 1654-1658 ◽  
Author(s):  
S Marcovina ◽  
D France ◽  
R A Phillips ◽  
S J Mao
Keyword(s):  
Monoclonal Antibodies ◽  
Apolipoprotein B ◽  
Polyclonal Antibodies ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Radial Immunodiffusion ◽  
Enzyme Linked Immunosorbent Assay ◽  
Low Density ◽  
Apo B ◽  
Blotting Technique

Abstract We produced 20 mouse monoclonal antibodies against human plasma low-density lipoprotein (LDL). Individually they failed to precipitate LDL in agarose gel by the double-immunodiffusion technique; collectively they did, or as few as two combined monoclonal antibodies could do so. To mimic polyclonal antibodies in determination of apolipoprotein B (apo B) by radial immunodiffusion, a combination of four particular monoclonal antibodies (clones A, B, C, and D) was necessary. We characterized these four clones with respect to temperature dependency, affinity, total binding to 125I-labeled LDL, and specificity to the different species of apolipoprotein B. Two monoclonal antibodies (B and C) bound 100% of 125I-labeled LDL; clones A and D bound 80% and 87%, respectively. All four clones bound maximally to LDL at 4 degrees C. The affinity constants for clones A, B, C, and D were 0.6, 2.1, 3.8, and 2.3 X 10(9) L/mol, respectively. By the Western blotting technique, the four monoclonal antibodies all reacted with the species B-100 and B-74 of apolipoprotein B, and to various degrees with B-48 and B-26. Radial immunodiffusion (chi) and direct enzyme-linked immunosorbent assay (y) with a mixture of the four monoclonal antibodies gave almost identical results for 70 patients: y = 0.921 chi-2.58; r = 0.933.

scholarly journals Application of Monoclonal Antibodies against Bioactive Natural Products: Eastern Blotting and Preparation of Knockout Extract

2012 ◽  
Vol 2012 ◽  
pp. 1-10 ◽  
Author(s):  
Hiroyuki Tanaka ◽  
Osamu Morinaga ◽  
Takuhiro Uto ◽  
Shunsuke Fuji ◽  
Frederick Asare Aboagye ◽  
...  
Keyword(s):  
No Production ◽  
Laser Desorption Ionization ◽  
Double Staining ◽  
Immunoaffinity Column ◽  
New Techniques ◽  
Pvdf Membrane ◽  
Bioactive Natural Products ◽  
Eastern Blotting ◽  
Complete Identification ◽  
Blotting Technique

Matrix-assisted laser desorption/ionization (MALDI) tof mass spectrometry was used for the confirmation of hapten number in synthesized antigen. As application of MAb, the MAbs against ginsenosides and glycyrrhizin have been prepared resulting in the development of two new techniques that we named the eastern blotting method and the knockout extract preparation. In eastern blotting technique, glycosides like ginsenosides and glycyrrhizin separated by silica gel TLC were blotted to PVDF membrane that was treated with a NaIO4solution followed by BSA resulted in glycoside-BSA conjugate on a PVDF membrane. The blotted spots were stained by MAb. Double staining of eastern blotting for ginsenosides using antiginsenoside Rb1and Rg1MAbs promoted complete identification of ginsenosides inPanaxspecies. The immunoaffinity concentration of glycyrrhizin was determined by immunoaffinity column conjugated with antiglycyrrhizin MAb resulting in the glycyrrhizin-knockout extract, which was determined by the synergic effect with glycyrrhizin on NO production using the cell line.

scholarly journals Development of Eastern Blotting Technique for Analysis of Baicalin Using Anti-Baicalin Monoclonal Antibody

Antibodies ◽  
2012 ◽  
Vol 1 (3) ◽  
pp. 284-293 ◽  
Author(s):  
Osamu Morinaga ◽  
Ryo Mukae ◽  
Takuhiro Uto ◽  
Yothawathorn Pariyawongsakul ◽  
Waraporn Putalun ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Eastern Blotting ◽  
Blotting Technique

scholarly journals Hybridomas for production of monoclonal antibodies to human erythropoietin

Blood ◽  
1984 ◽  
Vol 64 (2) ◽  
pp. 357-364 ◽  
Author(s):  
S Yanagawa ◽  
S Yokoyama ◽  
K Hirade ◽  
R Sasaki ◽  
H Chiba ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Growth Rate ◽  
Monoclonal Antibodies ◽  
Mechanism Of Action ◽  
Rapid Growth ◽  
Human Erythropoietin ◽  
Rapid Growth Rate ◽  
Blotting Technique ◽  
Purification Methods ◽  
High Production

Abstract Human urinary erythropoietin has been highly purified by a combination of conventional purification methods and immunoadsorbent columns packed with hybridoma-produced antibodies against contaminants that seemed difficult to separate from erythropoietin by the usual means. By using the partially purified erythropoietin as an antigen, three hybridoma clones have been obtained that secrete monoclonal antibodies against erythropoietin. One of the clones has been quite stable, with a rapid growth rate and high production of antibody. Western blotting technique with monoclonal antibodies revealed occurrence of two species of erythropoietin. The monoclonal antibody will be useful as a probe for the purification of erythropoietin and for further studies of the hormone and its mechanism of action.

Evaluation of a new eastern blotting technique for the analysis of ginsenoside Re in American ginseng berry pulp extracts

Fitoterapia ◽  
2010 ◽  
Vol 81 (4) ◽  
pp. 284-288 ◽  
Author(s):  
Osamu Morinaga ◽  
Takuhiro Uto ◽  
Chun-Su Yuan ◽  
Hiroyuki Tanaka ◽  
Yukihiro Shoyama
Keyword(s):  
American Ginseng ◽  
Ginsenoside Re ◽  
Eastern Blotting ◽  
Blotting Technique ◽  
Analysis Of Ginsenoside

scholarly journals Studies on Natural Products Using Monoclonal Antibodies: A Review

Antibodies ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 43
Author(s):  
Yukihiro Shoyama
Keyword(s):  
Natural Products ◽  
Monoclonal Antibodies ◽  
Thin Layer ◽  
Small Molecule ◽  
Polyvinylidene Fluoride ◽  
Herbal Medicines ◽  
Sugar Moiety ◽  
Eastern Blotting ◽  
Tlc Plates ◽  
Layer Chromatography

An immunoblotting system (“eastern blotting”) was developed for small-molecule herbal medicines like glycosides, with no conjugation function to the membrane. Briefly, the crude extracts of herb medicines were developed by thin-layer chromatography (TLC). The small-molecule herbal medicines on TLC plates were transferred to polyvinylidene fluoride (PVDF) or polyethersulfone (PES) membranes by heating. Antigen components were divided into two categories based on their function, i.e., their membrane recognizing (aglycone part) and fixing (sugar moiety) abilities. This procedure allows for the staining of only target glycosides. Double eastern blotting was developed as a further staining system for two herb medicines using a set of MAbs and substrates.

scholarly journals Hybridomas for production of monoclonal antibodies to human erythropoietin

Blood ◽  
1984 ◽  
Vol 64 (2) ◽  
pp. 357-364 ◽  
Author(s):  
S Yanagawa ◽  
S Yokoyama ◽  
K Hirade ◽  
R Sasaki ◽  
H Chiba ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Growth Rate ◽  
Monoclonal Antibodies ◽  
Mechanism Of Action ◽  
Rapid Growth ◽  
Human Erythropoietin ◽  
Rapid Growth Rate ◽  
Blotting Technique ◽  
Purification Methods ◽  
High Production

Human urinary erythropoietin has been highly purified by a combination of conventional purification methods and immunoadsorbent columns packed with hybridoma-produced antibodies against contaminants that seemed difficult to separate from erythropoietin by the usual means. By using the partially purified erythropoietin as an antigen, three hybridoma clones have been obtained that secrete monoclonal antibodies against erythropoietin. One of the clones has been quite stable, with a rapid growth rate and high production of antibody. Western blotting technique with monoclonal antibodies revealed occurrence of two species of erythropoietin. The monoclonal antibody will be useful as a probe for the purification of erythropoietin and for further studies of the hormone and its mechanism of action.

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