scholarly journals Double Staining of Ginsenosides by Western Blotting Using Anti-ginsenoside Rb1 and Rg1 Monoclonal Antibodies.

2001 ◽  
Vol 24 (10) ◽  
pp. 1157-1160 ◽  
Author(s):  
Noriko FUKUDA ◽  
Hiroyuki TANAKA ◽  
Yukihiro SHOYAMA
Keyword(s):  
Monoclonal Antibodies ◽  
Western Blotting ◽  
Ginsenoside Rb1 ◽  
Double Staining

scholarly journals Fingerprinting of Natural Product by Eastern Blotting Using Monoclonal Antibodies

2012 ◽  
Vol 2012 ◽  
pp. 1-7
Author(s):  
Hiroyuki Tanaka ◽  
Waraporn Putalun ◽  
Yukihiro Shoyama
Keyword(s):  
Monoclonal Antibodies ◽  
Natural Product ◽  
Cross Reactivity ◽  
Blue Color ◽  
Ginsenoside Rb1 ◽  
Double Staining ◽  
Eastern Blotting ◽  
The Central Nervous System ◽  
Rf Values ◽  
Solasodine Glycosides

We succeeded in developing the fingerprint of natural product by eastern blotting using monoclonal antibodies. After developing and separating them on a TLC plate, solasodine glycosides are oxidized by NaIO4 and reacted with a protein to give conjugates which are recognized with anti-solamargine monoclonal antibody (MAb). Anti-solamargine MAb having wide cross-reactivity can stain and detect all solasodine glycosides by fingerprint. Different sensitivity between solamargine and solasonine was observed. The detection limit was 1.6 ng of solasonine. The hydrolysed products of solamargine were determined by fingerprint of eastern blotting compared to their Rf values depending on the sugar number. Fingerprint by eastern blotting using anti-ginsenoside Rb1 MAb distinguished the formula containing ginseng prescribed in traditional Chinese medicine. By double-staining of ginsenosides it is possible to suggest that the staining color shows the pharmacological activity, such as the purple bands indicate ginsenosides having stimulation activity, and the blue color indicated compound like ginsenosides possessed the depression affect for the central nervous system (CNS), respectively.

Cross-Linked αsγt-Chain Hybrids in Plasma Clots Studied by 1D- and 2D Electrophoresis and Western Blotting

1993 ◽  
Vol 70 (03) ◽  
pp. 438-442 ◽  
Author(s):  
B Grøn ◽  
C Filion-Myklebust ◽  
S Bjørnsen ◽  
P Haidaris ◽  
F Brosstad
Keyword(s):  
Molecular Weight ◽  
Monoclonal Antibodies ◽  
Human Plasma ◽  
Western Blotting ◽  
Isoelectric Point ◽  
Factor Xiii ◽  
Cross Linking ◽  
2D Electrophoresis ◽  
Complex Phenomenon ◽  
Fibrinopeptide A

SummaryFibrinogen and fibrin related chains in reduced human plasma as well as the bonds interlinking partially cross-linked fibrin from plasma clots have been studied by means of 1D- and 2D electrophoresis and Western blotting. Immunovisualization of reduced plasma or partially cross-linked fibrin with monoclonal antibodies specific for the α-chains or the γ-chains have shown that several bands represent material belonging to both chains. In order to decide whether these bands constitute αγ-chain hybrids or superimposed α- and γ-chain dimers, the cross-linked material was separated according to both isoelectric point (pI) and molecular weight (MW) using Pharmacia’s Multiphor II system. Western blotting of the second dimension gels revealed that partially cross-linked fibrin contains αsγt-chain hybrids and γ- polymers, in addition to the well-known γ-dimers and α-polymers. The main αsγt-chain hybrid has a pI between that of the α- and the γ-chains, a MW of about 200 kDa and contains Aα-chains with intact fibrinopeptide A (FPA). It was also observed that soluble fibrinogen/fibrin complexes as well as partially cross-linked fibrin contain degraded α-dimers with MWs close to the γ-dimers. These findings demonstrate that factor XIII-catalyzed cross-linking of fibrin is a more complex phenomenon than earlier recognized.

scholarly journals Development of a New Highly Selective Monoclonal Antibody against Preferentially Expressed Antigen in Melanoma (PRAME) and Identification of the Target Epitope by Bio-Layer Interferometry

2021 ◽  
Vol 22 (6) ◽  
pp. 3166
Author(s):  
Jwala Priyadarsini Sivaccumar ◽  
Antonio Leonardi ◽  
Emanuela Iaccarino ◽  
Giusy Corvino ◽  
Luca Sanguigno ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Western Blotting ◽  
Cancer Biomarkers ◽  
Trypsin Digestion ◽  
Protein Fragments ◽  
Target Epitope ◽  
Potential Activity ◽  
Antibody Epitopes

Background: Monoclonal antibodies (mAbs) against cancer biomarkers are key reagents in diagnosis and therapy. One such relevant biomarker is a preferentially expressed antigen in melanoma (PRAME) that is selectively expressed in many tumors. Knowing mAb’s epitope is of utmost importance for understanding the potential activity and therapeutic prospective of the reagents. Methods: We generated a mAb against PRAME immunizing mice with PRAME fragment 161–415; the affinity of the antibody for the protein was evaluated by ELISA and SPR, and its ability to detect the protein in cells was probed by cytofluorimetry and Western blotting experiments. The antibody epitope was identified immobilizing the mAb on bio-layer interferometry (BLI) sensor chip, capturing protein fragments obtained following trypsin digestion and performing mass spectrometry analyses. Results: A mAb against PRAME with an affinity of 35 pM was obtained and characterized. Its epitope on PRAME was localized on residues 202–212, taking advantage of the low volumes and lack of fluidics underlying the BLI settings. Conclusions: The new anti-PRAME mAb recognizes the folded protein on the surface of cell membranes suggesting that the antibody’s epitope is well exposed. BLI sensor chips can be used to identify antibody epitopes.

Use of Commercially Available Rabbit Monoclonal Antibodies for Immunofluorescence Double Staining

2008 ◽  
Vol 16 (4) ◽  
pp. 387-392 ◽  
Author(s):  
Michael Bzorek ◽  
Inger Merete Stamp ◽  
Bodil Laub Petersen ◽  
Lisbeth Frederiksen
Keyword(s):  
Monoclonal Antibodies ◽  
Double Staining

Cellular phenotypes of normal and leukemic hemopoietic cells determined by analysis with selected antibody combinations

Blood ◽  
1980 ◽  
Vol 56 (3) ◽  
pp. 430-441 ◽  
Author(s):  
G Janossy ◽  
FJ Bollum ◽  
KF Bradstock ◽  
J Ashley
Keyword(s):  
Bone Marrow ◽  
Monoclonal Antibodies ◽  
Single Cell ◽  
Normal Cell ◽  
Cell Types ◽  
Precursor Cells ◽  
Leukemic Cells ◽  
Double Staining ◽  
Cell Assays ◽  
Cellular Phenotypes

Abstract Individual leukemic cells and the corresponding rare normal cell types in nonleukemic bone marrow were analyzed with various combinations of antisera (labeled with different fluorochromes: TRITC and FITC). Double staining for membrane Ia-like molecules (TRITC) and nuclear terminal transferase (FITC) was a very useful combination that distinguished common non-T, non-B ALL (Ia+,TdT+) and thymic ALL (Ia-,TdT+) from the rare cases of B ALL (Ia+,TdT-) and from AML (frequently Ia+, TdT-; in some cases Ia-, TdT-). Additional antisera (such as anti-ALL, anti- HuTLA, anti-immunoglobulin reagents, etc.) confirmed the diagnosis and further characterized the leukemic blasts. Ia+,TdT+ cells could be observed in low numbers in normal and nonleukemic regenerating marrow and were probably normal precursor cells; this reagent combinations was, therefore, not useful for monitoring residual non-T, non-B ALL blasts in treated patients. Other marker combinations detecting pre-B ALL blasts (double staining for cytoplasmic IgM and nuclear TdT) and Thy-ALL blasts (HuTLA+,TdT+) were, however, virtually leukemia specific in the bone marrow and could be used to effectively monitor residual leukemic cells throughout the disease. These combined single-cell assays are not only economical and informative but are also important for assessing the heterogeneity of leukemia and for standardizing new mouse or rat monoclonal antibodies for diagnosis.

A rapid and simple hapten conjugation method for monoclonal antibodies to be used in immunoenzyme single and double staining procedures

1987 ◽  
Vol 99 (2) ◽  
pp. 199-204 ◽  
Author(s):  
H. Mullink ◽  
D.M. Boorsma ◽  
J.C. Klein ◽  
R. Oostendorp ◽  
S.C. Henzen-Logmans ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Double Staining ◽  
Conjugation Method

Simultaneous detection of two independent antigens by double staining with two mouse monoclonal antibodies

1998 ◽  
Vol 73 (1) ◽  
pp. 89-97 ◽  
Author(s):  
Norihiro Teramoto ◽  
Laszlo Szekely ◽  
Katja Pokrovskaja ◽  
Li Fu Hu ◽  
Tadashi Yoshino ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Simultaneous Detection ◽  
Double Staining

scholarly journals Development, Characterization, and Application of Monoclonal Antibodies against Severe Acute Respiratory Syndrome Coronavirus Nucleocapsid Protein

2010 ◽  
Vol 17 (12) ◽  
pp. 2033-2036 ◽  
Author(s):  
Dipankar Das ◽  
Sriram Kammila ◽  
Mavanur R. Suresh
Keyword(s):  
Monoclonal Antibodies ◽  
Severe Acute Respiratory Syndrome ◽  
Western Blotting ◽  
Nucleocapsid Protein ◽  
Epitope Mapping ◽  
High Specificity ◽  
Diagnostic Assays ◽  
Hybridoma Technology ◽  
Recombinant Nucleocapsid Protein

ABSTRACT Five monoclonal antibodies (MAbs) against recombinant nucleocapsid protein (NP) of severe acute respiratory syndrome (SARS)-causing coronavirus (CoV) were developed by hybridoma technology. Epitope mapping by Western blotting showed that these anti-SARS-CoV NP MAbs bind to distinct domains of NP. These anti-SARS-CoV NP MAbs, with their high specificity, are potentially ideal candidates for developing early and sensitive diagnostic assays for SARS-CoV.

Sign in / Sign up

Export Citation Format

Share Document