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1.
James R. Wilson Milton M. Weiser Boris Albini Jay R. Schenck Harry G. Rittenhouse A.A. Hirata Eric G. Berger 《Biochemical and biophysical research communications》1982,105(2):737-744
Preparations of human malignant effusion galactosyltransferase activity purified according to previously published techniques using enzyme-specific affinity chromatography consistently produced antibodies directed toward immunoglobulins with no detectable antigalactosyltransferase. Double immunodiffusion analysis of the antigen showed the presence of both IgG and IgA. Affinity chromatography with anti-human IgG-Sepharose and anti-human serum-Sepharose resulted in a 48,000-fold purification of galactosyltransferase activity with no detectable IgG by radioimmunoassay. Immunization of rabbits with this preparation produced antibodies directed against galactosyltransferase activity and minimal anti-Ig. The persistence of immunoglobulins during the purification of soluble galactosyltransferase activity through two enzyme-specific affinity chromatographic steps suggests an association of immunoglobulins with galactosyltransferase activity. 相似文献
2.
The chemiluminescence immunoassay for aflatoxin B1 based on functionalized magnetic nanoparticles with two strategies of antigen probe immobilization 下载免费PDF全文
A rapid and sensitive chemiluminescence immunoassay (CLIA) based on magnetic nanoparticles (MNPs) was developed to detect aflatoxin B1 (AFB1), which is a potent carcinogen in nature. We prepared monodisperse MNPs (300 nm in diameter) according to the solvothermal synthesis reaction and the MNPs were coated with silica by the Stöber method. Triethox was used as a one‐step carboxylation reagent, and 3‐aminopropyltriethoxysilane (APTES) an amination reagent, to modify the MNPs. We prepared two types of solid phase antigens using the above synthesized functionalized MNPs coupled with the later prepared AFB1‐oxime active ester and the purchased BSA–AFB1 respectively. 2′,6′‐dimethylcarbonylphenyl‐10‐sulfopropylacridinium‐9‐carboxylate 4′‐N‐hydroxysuccinimide (4′‐NHS) ester (NSP–DMAE–NHS), as a novel luminescent reagent, was used to label anti‐AFB1 antibodies. The two CLIA calibration curves based on the two types of solid phase antigens were obtained and compared. The acquired limit of detection (LOD) was about 0.001 ng/mL for the two functionalized MNPs‐based immunoassays, and the half maximal inhibitory concentration (IC50) was 0.51 ng/mL for the MNPs–AFB1‐based method and 0.72 ng/mL for the MNPs–BSA–AFB1‐based method. 相似文献
3.
Nance-Horan Syndrome (NHS) is a rare X-linked syndrome characterized by congenital cataract which leads to profound vision loss, characteristic dysmorphic features and specific dental anomalies. Microcornea, microphthalmia and mild or moderate mental retardation may accompany these features. Heterozygous females often manifest similarly but with less severe features than affected males. We describe two brothers who have the NHS phenotype and their carrier mother who had microcornea but not cataract. We identified a previously unreported frameshift mutation (c.558insA) in exon 1 of the NHS gene in these patients and their mother which is predicted to result in the incorporation of 11 aberrant amino acids prior to a stop codon (p.E186Efs11X). We also discussed genotype–phenotype correlation according to relevant literature. 相似文献
4.
Xiaohui Liang Xiangyi Fang Manwen Yao Yucong Yang Junfeng Li Hongjun Liu Linyu Wang 《Luminescence》2016,31(1):168-172
Direct competitive chemiluminescence immunoassays (CLIA) based on gold‐coated magnetic nanospheres (Au‐MNPs) were developed for rapid analysis of chloramphenicol (CAP). The Au‐MNPs were modified with carboxyl groups and amino groups by 11‐mercaptoundecanoic acid (MUA) and cysteamine respectively, and then were respectively conjugated with CAP base and CAP succinate via an activating reaction using 1‐ethyl‐3‐(3‐dimethylaminopropyl)carbodiimide hydrochloride (EDC) and N‐hydroxysuccinimide (NHS). NSP‐DMAE‐NHS, a new and effective luminescence reagent, was employed to label anti‐CAP antibody (mAb) as a tracer in direct CLIA for CAP detection using a ‘homemade’ luminescent measurement system that was set up with a photomultiplier tube (PMT) and a photon counting unit linked to a computer. The sensitivities and limits of detection (LODs) of the two methods were obtained and compared according to the inhibition curves. The 50% inhibition concentration (IC50) values of the two methods were about 0.044 ng/mL and 0.072 ng/mL respectively and LODs were approximately 0.001 ng/mL and 0.006 ng/mL respectively. To our knowledge, they were much more sensitive than any traditional enzyme‐linked immunosorbent assay (ELISA) ever reported. Moreover, the new luminescence reagent NSP‐DMAE‐NHS is much more sensitive and stable than luminol and its derivatives, contributing to the sensitivity enhancement. Copyright © 2015 John Wiley & Sons, Ltd. 相似文献
5.
Kazuhiro Hasegawa Daisaku OzawaTadakazu Ookoshi Hironobu Naiki 《Biochimica et Biophysica Acta - Proteins and Proteomics》2013,1834(8):1624-1631
Cerebral amyloid angiopathy is caused by deposition of the amyloid β-peptide which consists of mainly 39–40 residues to the cortical and leptomeningeal vessel walls. There are no definite in vitro systems to support the hypothesis that the vascular basement membrane may act as a scaffold of amyloid β-peptide carried by perivascular drainage flow and accelerate its amyloid fibril formation in vivo. We previously reported the critical roles of interfaces and agitation on the nucleation of amyloid fibrils at low concentrations of amyloid β-peptide monomers. Here, we reproduced the perivascular drainage flow in vitro by using N-hydroxysuccinimide-Sepharose 4 Fast flow beads as an inert stirrer in air-free wells rotated at 1 rpm. We then reproduced the basement membranes in the media of cerebral arteries in vitro by conjugating Matrigel and other proteins on the surface of Sepharose beads. These beads were incubated with 5 μM amyloid β(1–40) at 37 °C without air, where amyloid β(1–40) alone does not form amyloid fibrils. Using the initiation time of fibril growth kinetics (i.e., the lag time of fibril growth during which nuclei, on-pathway oligomers and protofibrils are successively formed) as a parameter of the efficiency of biological molecules to induce amyloid fibril formation, we found that basement membrane components including Matrigel, laminin, fibronectin, collagen type IV and fibrinogen accelerate the initiation of amyloid β-peptide fibril growth in vitro. These data support the essential role of vascular basement membranes in the development of cerebral amyloid angiopathy. 相似文献
6.
Seong-Cheol Park Il Ryong Kim Jin-Young Kim Yongjae Lee Eun-Ji Kim Ji Hyun Jung Young Jun Jung Mi-Kyeong Jang Jung Ro Lee 《Biochimica et Biophysica Acta (BBA)/General Subjects》2018,1862(12):2545-2554
Background
It remains an open question whether plant phloem sap proteins are functionally involved in plant defense mechanisms.Methods
The antifungal effects of two profilin proteins from Arabidopsis thaliana, AtPFN1 and AtPFN2, were tested against 11 molds and 4 yeast fungal strains. Fluorescence profiling, biophysical, and biochemical analyses were employed to investigate their antifungal mechanism.Results
Recombinant AtPFN1 and AtPFN2 proteins, expressed in Escherichia coli, inhibited the cell growth of various pathogenic fungal strains at concentrations ranging from 10 to 160?μg/mL. The proteins showed significant intracellular accumulation and cell-binding affinity for fungal cells. Interestingly, the AtPFN proteins could penetrate the fungal cell wall and membrane and act as inhibitors of fungal growth via generation of cellular reactive oxygen species and mitochondrial superoxide. This triggered the AtPFN variant-induced cell apoptosis, resulting in morphological changes in the cells.Conclusion
PFNs may play a critical role as antifungal proteins in the Arabidopsis defense system against fungal pathogen attacks.General significance
The present study indicates that two profilin proteins, AtPFN1 and AtPFN2, can act as natural antimicrobial agents in the plant defense system. 相似文献7.
G. Mattson E. Conklin S. Desai G. Nielander M. D. Savage S. Morgensen 《Molecular biology reports》1993,17(3):167-183
The various aspects of chemical crosslinking are addressed. Crosslinker reactivity, specificity, spacer arm length and solubility characteristics are detailed. Considerations for choosing one of these crosslinkers for a particular application are given as well as reaction conditions and practical tips for use of each category of crosslinkers.Abbreviations ABH
azidobenzoyl hydrazide
- ANB- NOS
N-5-azido-2-nitrobenzoyloxysuccinimide
- ASIB
1-(p-azidosalicylamido)-4-(iodoacetamido)butane
- ASBA
4-(p-azidosalicylamido)butylamine
- APDP
N-[4-(p-azidosalicylamido) butyl]-3(2-pyridyldithio)propionamide
- APG
p-azidophenyl glyoxal monohydrate
- BASED
bis-[-(4-azidosalicylamido)ethyl] disulfide
- BMH
bismaleimidohexane
- BS3
bis(sulfosuccinimidyl) suberate
- BSOCOES
bis[2-(succinimidooxycarbonyloxy)ethyl]sulfone
- DCC
N,N-dicyclohexylcarbodiimide
- DFDNB
1,5-difluoro-2,4-dinitrobenzene
- DMA
dimethyl adipimidate·2HCl
- DMP
dimethyl pimelimidate·2HCl
- DMS
dimethyl suberimidate·2HCl
- DPDPB
1,4-di-(3,2-pyridyldithio)propionamido butane
- DMF
dimethylformamide
- DMSO
dimethylsulfoxide
- DSG
disuccinimidyl glutarate
- DSP
dithiobis(succinimidylpropionate)
- DSS
disuccinimidyl suberate
- DST
disuccinimidyl tartarate
- DTSSP
3,3-dithiobis (sulfosuccinimidylpropionate)
- DTBP
dimethyl 3,3-dithiobispropionimidate·2HCl
- EDC or EDAC
1-ethyl-3-(3-dimethylaminopropyl)carbodimide hydrochloride
- EDTA
ethylenediaminetetraacetic acid disodium salt, dihydrate
- EGS
ethylene glycolbis(succinimidylsuccinate)
- GMBS
N--maleimidobutyryloxysuccinimide ester
- HSAB
N-hydroxysuccinimidyl-4-azidobenzoate
- HEPES
4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid
- MBS
m-maleimidobenzoyl-N-hydroxysuccinimide ester
- MES
4-morpholineethanesulfonic acid
- NHS
N-hydroxysuccinimide
- NHS-ASA
N-hydroxysuccinimidyl-4-azidosalicylic acid
- PMFS
phenylmethylsulfonyl fluoride
- PNP-DTP
p-nitrophenyl-2-diazo-3,3,3-trifluoropropionate
- SAED
sulfosuccinimidyl 2-(7-azido-4-methylcoumarin-3-acetamide) ethyl-1,3-dithiopropionate
- SADP
N-succinimdyl (4-azidophenyl)1,3-dithiopropionate
- SAND
sulfosuccinimidyl 2-(m-azido-o-nitrobenzamido)-ethyl-1,3-dithiopropionate
- SANPAH
N-succinimidyl-6(4-azido-2-nitrophenyl-amino)hexanoate
- SASD
sulfosuccinimidyl 2-(p-azidosalicylamido)ethyl-1,3-dithiopropionate
- SATA
N-succinimidyl-S-acetylthioacetate
- SDBP
N-hydroxysuccinimidyl-2,3-dibromopropionate
- SIAB
N-succinimidyl(4-iodoacetyl)aminobenzoate
- SMCC
succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate
- SMPB
succinimidyl 4-(p-maleimidophenyl) butyrate
- SMPT
4-succinimidyloxycarbonyl--methyl--(2-pyridyldithio)-toluene
- sulfo-BSOCOES
bis[2-sulfosuccinimidooxycarbonyloxy) ethyl]sulfone
- sulfo-DST
disulfosuccinimidyl tartarate
- sulfo-EGS
ethylene glycolbis(sulfosuccinimidylsuccinate)
- sulfo-GMBS
N--maleimidobutyryloxysulfosuccinimide ester
- sulfo-MBS
m-maleimidobenzoyl-N-hydroxysulfosuccinimide ester
- sulfo-SADP
sulfosuccinimidyl(4-azidophenyldithio)propionate
- sulfo-SAMCA
sulfosuccinimidyl 7-azido-4-methylcoumarin-3-acetate
- sulfo-SANPAH
sulfosuccinimidyl 6-(4-azido-2-nitrophenylamino)hexanoate
- sulfo-SIAB
sulfosuccinimidyl(4-iodoacetyl)aminobenzoate
- sulfo-SMPB
sulfo-succinimidyl 4-(p-maleimidophenyl)butyrate
- sulfo-SMCC
sulfosuccinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate
- SPDP
N-succinimidyl 3-(2-pyridyldithio)propionate 相似文献
8.
Kacee H. Sims Ewan M. Tytler John Tipton Kasey L. Hill Stephen W. Burgess Walter A. Shaw 《Biochimica et Biophysica Acta (BBA)/Molecular and Cell Biology of Lipids》2014,1841(8):1038-1048
Lipid research is challenging owing to the complexity and diversity of the lipidome. Here we review a set of experimental tools developed for the seasoned lipid researcher, as well as, those who are new to the field of lipid research. Novel tools for probing protein–lipid interactions, applications for lipid binding antibodies, enhanced systems for the cellular delivery of lipids, improved visualization of lipid membranes using gold-labeled lipids, and advances in mass spectrometric analysis techniques will be discussed. Because lipid mediators are known to participate in a host of signal transduction and trafficking pathways within the cell, a comprehensive lipid toolbox that aids the science of lipidomics research is essential to better understand the molecular mechanisms of interactions between cellular components. This article is part of a Special Issue entitled Tools to study lipid functions. 相似文献
9.
Marion Babot Paola Labarbuta Amanda Birch Sara Kee Matthew Fuszard Catherine H. Botting Ilka Wittig Heinrich Heide Alexander Galkin 《BBA》2014
An intriguing feature of mitochondrial complex I from several species is the so-called A/D transition, whereby the idle enzyme spontaneously converts from the active (A) form to the de-active (D) form. The A/D transition plays an important role in tissue response to the lack of oxygen and hypoxic deactivation of the enzyme is one of the key regulatory events that occur in mitochondria during ischaemia. We demonstrate for the first time that the A/D conformational change of complex I does not affect the macromolecular organisation of supercomplexes in vitro as revealed by two types of native electrophoresis. Cysteine 39 of the mitochondrially-encoded ND3 subunit is known to become exposed upon de-activation. Here we show that even if complex I is a constituent of the I + III2 + IV (S1) supercomplex, cysteine 39 is accessible for chemical modification in only the D-form. Using lysine-specific fluorescent labelling and a DIGE-like approach we further identified two new subunits involved in structural rearrangements during the A/D transition: ND1 (MT-ND1) and 39 kDa (NDUFA9). These results clearly show that structural rearrangements during de-activation of complex I include several subunits located at the junction between hydrophilic and hydrophobic domains, in the region of the quinone binding site. De-activation of mitochondrial complex I results in concerted structural rearrangement of membrane subunits which leads to the disruption of the sealed quinone chamber required for catalytic turnover. 相似文献
10.
Majumdar S Burgman M Haselton N Grinnell S Ocampo J Pasternak AR Pasternak GW 《Bioorganic & medicinal chemistry letters》2011,21(13):4001-4004
Tritiated opioid radioligands have proven valuable in exploring opioid binding sites. However, tritium has many limitations. Its low specific activity and limited counting efficiency makes it difficult to examine low abundant, high affinity sites and its disposal is problematic due to the need to use organic scintillants and its relatively long half-life. To overcome these issues, we have synthesized both unlabeled and carrier-free radioiodinated iodobenzoyl derivatives of 6β-naltrexamine (125I-BNtxA, 18), 6β-naloxamine (125I-BNalA, 19) and 6β-oxymorphamine (125I-BOxyA, 20) with specific activities of 2100 Ci/mmol. To optimize the utility of the radioligand, we designed a synthesis in which the radiolabel is incorporated in the last synthetic step, which required the selective iodination of the benzoyl moiety without incorporation into the phenolic A ring. Competition studies demonstrated high affinity of the unlabelled compounds for opioid receptors in transfected cell lines, as did the direct binding of the 125I-ligands to the opioid receptors. The radioligand displayed very high sensitivity, enabling a marked reduction in tissue, as well as excellent signal/noise characteristics. These new 125I-radioligands should prove valuable in future studies of opioid binding sites. 相似文献