Abstract：
:The characterization of mucins is critically important for gaining insights into the molecular pathology of diseases, including cancers, as well as for the discovery of biomarkers for disease diagnosis. However, no practical method has yet been reported for identifying mucin proteins. Here, we report a technique for immunological identification of mucins separated by supported molecular matrix electrophoresis (SMME), a recently developed membrane electrophoresis method. The technique involves on-membrane deglycosylation of mucins by using mild periodate oxidation/base-catalyzed elimination, followed by immunostaining with an antibody that specifically recognizes the mucin tandem repeat (TR) peptide. We demonstrated the method's feasibility by using MUC1 derived from 2 cancer cell lines, T47D and HPAF-II. The present method shows potential as an alternative approach for the identification of mucins separated by SMME or blotted from conventional gel electrophoresis, on a PVDF membrane.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Matsuno YK,Dong W,Yokoyama S,Yonezawa S,Narimatsu H,Kameyama Adoi
10.1016/j.jim.2013.06.002subject
Has Abstractpub_date
2013-08-30 00:00:00pages
125-30issue
1-2eissn
0022-1759issn
1872-7905pii
S0022-1759(13)00174-9journal_volume
394pub_type
杂志文章abstract：:Rheumatoid peripheral blood mononuclear cells show an increased responsiveness to superinfection with Epstein-Barr virus (EBV). We have investigated the role of adherent cells in this hyperresponsiveness using two different methods of adherent cell depletion. Depletion of adherent cells from both rheumatoid and normal...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(87)90463-7
更新日期：1987-03-12 00:00:00
abstract：:We present a scheme for routine screening of patients' sera for monoclonal immunoglobulin by isoelectric focusing in agarose gels. Using this method combined with immunofixation, we have been able to detect monoclonal immunoglobulin in the sera of patients, and to identify the isotype of the immunoglobulin when conven...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90393-9
更新日期：1983-11-11 00:00:00
abstract：:Certain serum samples produce high background in Western and direct immunoblot assays that detect human serum IgG against specific antigens. We determined that this was due to a reaction between endogenous IgG and the membrane blocking agent (we refer to this as blocking-specific background). Using milk as blocking ag...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(93)90259-a
更新日期：1993-01-14 00:00:00
abstract：:Development of cell lines for biotherapeutic protein production requires screening large numbers of clones to identify and isolate high producing ones. As such, stable cell line generation is a time- and resource-intensive process. There is an increasing need to enhance the selection efficiency of high-yielding clonal...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2018.10.007
更新日期：2019-01-01 00:00:00
abstract：:Phycobilisomes are cyanobacterial photosynthetic energy transfer complexes partly composed of phycobiliproteins, proteins that are widely used as conjugable fluorochromes for flow cytometry. The brightness and photostability of phycobiliproteins suggest that intact phycobilisomes could constitute even brighter probes ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(01)00367-2
更新日期：2001-08-01 00:00:00
abstract：:To date, phase I/II dendritic cell (DC)-based cancer vaccine trials have required repeated venesection or leukapheresis to generate the DCs. Previous studies have suggested that DCs may be cryopreserved and revived for clinical use as sequential immunisations. We have developed a method of cryopreserving monocyte-deri...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(02)00430-1
更新日期：2003-01-15 00:00:00
abstract：:The binding of cells to paramagnetic polystyrene microbeads in the absence of coupling antibodies was measured. Cells from normal bone marrow, from an acute lymphoblastic leukemia (ALL) cell line or from a neuroblastoma cell line, were labeled with the fluorochrome Hoechst 33342 and incubated with microbeads by rotati...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(89)90173-7
更新日期：1989-07-26 00:00:00
abstract：:The study of organogenesis, tissue-homeostasis and regeneration requires the precise assessment of in vivo cell proliferation. To this end a host of methods have been developed to detect and quantify DNA synthesis in proliferating cells. These include cell labeling with various nucleotide analogues and fluorescence re...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2018.03.015
更新日期：2018-06-01 00:00:00
abstract：:Langerin CD207 is a type II transmembrane protein. It is responsible for the formation of Birbeck granules, which are intracellular organelles within Langerhans cells, the dendritic cells of stratified squamous epithelia like the epidermis. Because current anti-CD207 antibodies have limitations, we prepared new monocl...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2007.05.001
更新日期：2007-07-31 00:00:00
abstract：:Cow's milk protein allergy (CMPA) is one of the most prevalent human food-borne allergies, particularly in children. Experimental animal models have become critical tools with which to perform research on new therapeutic approaches and on the molecular mechanisms involved. However, oral food allergen sensitization in ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2012.04.007
更新日期：2012-07-31 00:00:00
abstract：INTRODUCTION:Basophils play a major physio-pathological role in hypersensitivity related diseases. Basophils express high affinity Immunoglobulin (Ig) E receptors (FcεRI), IgG and complement regulatory. Basophils also have immunoregulatory activity through interaction with T cells. The aim of this study was to look for...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2020.112951
更新日期：2021-01-22 00:00:00
abstract：:A fluorometric enzyme immunoassay using an alkaline phosphatase-conjugated monoclonal antibody was developed to quantitate feline leukemia virus (FeLV) infection. Monoclonal antibodies, directed against the FeLV structural protein p27, were conjugated with alkaline phosphatase using a modified maleimide method. The en...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(88)90181-0
更新日期：1988-11-10 00:00:00
abstract：:This study describes a four-color flow cytometric assay that detects CD4+ T cell responses to the anthrax vaccine. Whole blood from seven volunteers who previously obtained the anthrax vaccine was inoculated in vitro with varying concentrations of the anthrax antigen. TNF-alpha and IFN-gamma production from memory CD4...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2003.07.012
更新日期：2003-11-01 00:00:00
abstract：:Protein A containing staphylococci were saturated with human monomeric IgG (mIgG) and cross-linked with glutaraldehyde. The resulting material (SMG) preferentially bound aggregated IgG (aIgG) and soluble immune complexes (CIC). One milliliter of a 10% suspension of SMG bound approximately 30 micrograms of mIgG and 100...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(84)90259-x
更新日期：1984-01-20 00:00:00
abstract：:An enzyme-linked immunosorbent assay (ELISA) for the quantitation of rat erythrocyte membrane sialoglycoproteins (glycophorins) has been developed. Samples of erythrocytes and reticulocytes were analysed using this assay, and response compared among them and purified glycophorins samples. A broadly homologous behaviou...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(93)90387-m
更新日期：1993-06-18 00:00:00
abstract：:Radioactive double labeling was adapted for the analysis of mediators of cellular immunity. Two identical lymphocyte cultures were simultaneously labeled with [3H]- or [14C]leucine. Each of the cultures was stimulated with antigen or mitogen. The combined supernatants were then subjected to various fractionation proce...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(78)90177-1
更新日期：1978-01-01 00:00:00
abstract：:We describe a method for standardizing cytotoxicity assays by the use of cryopreserved, fluorescently labeled target cells. The cells are labeled in batches with the fluorogenic element europium (Eu) and frozen in multiple aliquots. Replicate aliquots can be thawed on different days and used for cytotoxicity assays. T...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(90)90112-9
更新日期：1990-11-06 00:00:00
abstract：:Investigation of MHC class I endogenous antigen processing and presentation is important for understanding non-self immune recognition, vaccine design, immune evasion by pathogens and tumours, and elucidation of the mechanisms of certain auto-immune diseases. To study the MHC class I antigen-processing pathway, the pr...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2008.02.017
更新日期：2008-06-01 00:00:00
abstract：:BlockLogo is a web-server application for the visualization of protein and nucleotide fragments, continuous protein sequence motifs, and discontinuous sequence motifs using calculation of block entropy from multiple sequence alignments. The user input consists of a multiple sequence alignment, selection of motif posit...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2013.08.014
更新日期：2013-12-31 00:00:00
abstract：:The method of extraction of outer membrane proteins (OMPs), the conditions of electrophoretic transfer, and the conditions of antibody binding, were compared in Western blotting studies of Neisseria meningitidis outer membrane antigens. The OMP profiles obtained by SDS-PAGE of outer membrane vesicles extracted with li...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(90)90383-7
更新日期：1990-12-05 00:00:00
abstract：:A plasmid cassette vector was designed to generate transgenic mice expressing reporter cDNAs at high levels in antigen-presenting cells under the control of the murine invariant chain (Ii) promoter. Analysis of several transgenic mice harboring a chimeric Ii cDNA placed in this vector showed that it can drive expressi...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(00)00276-3
更新日期：2000-11-01 00:00:00
abstract：:The fluorescence activated cell sorter (FACS) was used with an indirect membrane immunofluorescence technique to detect antibody against the Forssman antigen, a glycosphingolipid. Sheep erythrocytes, which contain Forssman antigen as a major membrane glycosphingolipid, were used as the target antigen. Detection of the...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90098-4
更新日期：1983-02-25 00:00:00
abstract：:The paper by Herzyk et al. which appears in this issue of the Journal of Immunological Methods is an important and long overdue contribution. The study clearly and convincingly demonstrates that two widely used commercial ELISA kits fail to detect over 90% of the 31 kDa human IL-1 beta precursor (pro-IL-1 beta) in var...
journal_title：Journal of immunological methods
pub_type： 评论,杂志文章
doi：10.1016/0022-1759(92)90179-w
更新日期：1992-04-08 00:00:00
abstract：:The aim of this study was to compare the techniques of indirect immunofluorescence assay (IFA) and flow cytometry to clinical and laboratorial evaluation of patients before and after clinical cure and to evaluate the applicability of flow cytometry in post-therapeutic monitoring of patients with American tegumentary l...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2012.11.002
更新日期：2013-01-31 00:00:00
abstract：:This paper describes a novel method for following the changes in mouse tumour-infiltrating immune cell populations by repeated sampling of tumours by fine needle aspiration (FNA), followed by flow cytometry. Using this technique we were able to collect samples from P815 mouse mastocytomas, and identify and enumerate s...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2015.06.015
更新日期：2015-10-01 00:00:00
abstract：:The sensitivity of any immunoassay is a complex function of the underlying physico-chemical basis of the technique and the size and source of 'experimental' errors. Analysis of this relationship emphasizes the distinction between competitive labelled analyte methods, e.g., RIA, and the non-competitive labelled antibod...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(86)90338-8
更新日期：1986-02-27 00:00:00
abstract：:Multivalent antibody fragments (scFv dimers, trimers and tetramers) provide high avidity and ideal pharmacokinetics for tumour targeting applications. This protocol describes our optimised protocol for high-level bacterial synthesis of soluble antibody scFv fragments, as either monomers or multimers, using the heat-in...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(00)00201-5
更新日期：2000-08-28 00:00:00
abstract：:A high non-specific binding of immunoglobulins to plastic surfaces was noted with a number of rat sera, when tested in an indirect 125I-labeled protein-A assay for detection of cell-surface-bound rat immunoglobulins of various classes and IgG subclasses. This type of non-specific binding was found with all types of Ig...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(84)90243-6
更新日期：1984-01-20 00:00:00
abstract：:The use of colloidal gold and colloidal gold followed by silver enhancement as marker for dot or blot immune overlays is described. Colloidal gold probes concentrating at the sites of immune reaction gradually develop a pinkish colour during incubation that can be seen with the naked eye. With a physical developer, ve...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(84)90303-x
更新日期：1984-11-30 00:00:00
abstract：:A one-step immunospecific affinity chromatographic method for purification of mouse alpha-fetoprotein (AFP) directly from amniotic fluid is described. The procedure employed a rabbit anti-AFP immunoglobulin-G entrapped in a polyacrylamide gel matrix prepared as a gel slurry. The presence of impurities could not be dem...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(79)90146-7
更新日期：1979-01-01 00:00:00
