A simplified solid-phase assay for the quantitation of native membrane proteins. Application to the measurement of EGF receptor induction by dexamethasone.

Abstract:

:Direct application of membrane fractions to a nitrocellulose support without previous solubilization, denaturation or fixation of the antigen permits the quantitation of antibodies binding to denaturation sensitive epitopes. In experiments reported here, this method was used to determine the rate of EGF receptor expression on squamous carcinoma cell lines. The assay revealed that dexamethasone treatment leads to an eight-fold increase of EGF receptor protein expression on C4-I cervical carcinoma cells. The data, when compared to the results obtained in immunoprecipitation experiments, suggest that this simple protocol yields reliable and precise quantitative data. The simplicity of the method permits simultaneous testing of large sample numbers with various antibodies.

journal_name

J Immunol Methods

authors

Von Knebel Doeberitz M,Drzonek H,Koch S,Becker CM

doi

10.1016/0022-1759(89)90272-x

subject

Has Abstract

pub_date

1989-09-01 00:00:00

pages

259-64

issue

2

eissn

0022-1759

issn

1872-7905

pii

0022-1759(89)90272-X

journal_volume

122

pub_type

杂志文章
  • Use of a radioactive double labeling technique in the chemical analysis of the mediators of cellular immunity.

    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

    authors: Sorg C

    更新日期:1978-01-01 00:00:00

  • Comparison of five calculation modes for antibody ELISA procedures using pertussis serology as a model.

    abstract::During a phase III pertussis vaccine trial, serum antibody responses were measured by two enzyme-linked immunosorbent assays (ELISA) for pertussis toxin and filamentous haemagglutinin. These were used both for studies of antibody levels after vaccination and for diagnostic purposes. Since the absorbance values obtaine...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(95)00067-k

    authors: Reizenstein E,Hallander HO,Blackwelder WC,Kühn I,Ljungman M,Möllby R

    更新日期:1995-06-28 00:00:00

  • A novel antibody overlay technique for two-dimensional immunoelectrophoresis.

    abstract::A new antibody overlay technique for 2-dimensional immunoelectrophoresis (2-D-IEP) is described. After the first dimension electrophoresis of the antigen, the desired amount of antiserum is applied to the initial agarose layer and then evenly distributed over a defined surface of the gel with a PVC film. This modifica...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(84)90332-6

    authors: Lebecq JC,Salhi SL,Bastide JM

    更新日期:1984-02-10 00:00:00

  • Immunoassays for inhibin and its subunits. Further applications of the synthetic peptide approach.

    abstract::We describe the preparation of a new rat monoclonal antibody (CRC1) to the N-terminal sequence of the 43 kDa subunit of human ovarian inhibin, and its use together with other anti-peptide monoclonal antibodies, in two-site immunoassays for the detection of inhibin-related material in biological fluids. The Fab fractio...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(93)90342-5

    authors: Groome N,O'Brien M

    更新日期:1993-10-15 00:00:00

  • Purification of uteroglobin using monospecific antibodies coupled to divinylsulphone-activated agarose.

    abstract::As a model for the isolation of a labile or trace protein, the purification of uteroglobin (UGL) by immunoaffinity chromatography is described. Antibody was isolated from sheep antiserum by immunoprecipitation, and coupled to divinylsulphone-activated agarose (Mini Leak). For the immunoabsorption stage rabbit uterine ...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(89)90400-6

    authors: Dannhorn DR,Wirth B,Kirchner C

    更新日期:1989-05-12 00:00:00

  • A mixed hemadsorption assay for detection of cell surface binding anti-tumor antibodies in human sera.

    abstract::A practical mixed hemadsorption assay (MHA) was developed for the detection of cell surface binding anti-tumor antigen antibodies (anti-TA Ab) in the human sera. The assay was compared to enzyme-linked immunosorbent assay (ELISA) and dot ELISA in the detection of the antibodies developed against GA733-2 antigen. SW111...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/s0022-1759(01)00560-9

    authors: Akis N,Andl C,Zhou D

    更新日期:2002-03-01 00:00:00

  • Discrimination by rabbit anti-idiotypic antibodies of two murine IgM monoclonal antibodies directed against lipid A.

    abstract::Two murine IgM monoclonal antibodies (MAs) directed against the lipid A portion of bacterial lipopolysaccharide (LPS) were compared in their binding to Re LPS and lipid A and their idiotypic make-up with rabbit anti-idiotypic sera. Horseradish peroxidase (HRPO)-labelled MAs 8-2 and 26-20 bound equally well to Re LPS. ...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(90)90308-i

    authors: Cornelissen JJ,Oosterlaken TA,Schellekens J,Torensma R,Rozenberg-Arska M,Kraaijeveld CA,Verhoef J

    更新日期:1990-06-12 00:00:00

  • Quantitation of messenger RNA by competitive RT-PCR: a simplified read out assay.

    abstract::A competitive RT-PCR method that permits reliable quantification of minute amounts of reverse-transcribed mouse lymph node mRNA is described. Using this technique, an absolute number of cDNA copies ranging from 10(3) to 10(5) can be determined, with a precision superior to 25%. The standard templates described in the ...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/s0022-1759(97)00186-5

    authors: Colle JH,Falanga PB,Singer M,Hevin B,Milon G

    更新日期:1997-12-29 00:00:00

  • Standardisation of a procedure for quantifying surface antigens by indirect immunofluorescence.

    abstract::Quantitative indirect immunofluorescence (QIIF) methods used to measure absolute numbers of surface-expressed antigens have produced conflicting results [Marchant, A., Duchow, J., Delville, J., Goldman, M., 1992. Lipopolysaccharide induces up-regulation of CD14 molecule on monocytes in human whole blood. European Jour...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/s0022-1759(99)00087-3

    authors: Smith KB,Ellis SA

    更新日期:1999-08-31 00:00:00

  • Introduction of a selectable gene into murine T-lymphoblasts by a retroviral vector.

    abstract::A recombinant retroviral vector with an inserted bacterial neomycin resistance (neo) gene was used to transfer in vitro neomycin resistance (neoR) into murine cytotoxic lymphocyte precursors (CLP). The infection protocol involved co-cultivation of mitogen-activated splenic T-blasts with irradiated cells that produced ...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(86)90036-0

    authors: Reimann J,Heeg K,Wagner H,Keller G,Wagner EF

    更新日期:1986-05-01 00:00:00

  • A flow cytometric rosetting assay for the analysis of IgG-Fc receptor interactions.

    abstract::We have developed a sensitive and flexible method for the qualitative evaluation of IgG-Fc receptor interactions in cell suspensions. The assay is based on the flow cytometric quantitation of antibody-coated erythrocyte (EA) rosetting using fluorescein-labelled indicator erythrocytes (E). The number of IgG molecules o...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(90)90070-c

    authors: Tuijnman WB,Van de Winkel JG,Capel PJ

    更新日期:1990-03-09 00:00:00

  • Automated complement fixation test for the detection of antibodies against the core of hepatitis B virus (HBc).

    abstract::The automated complement fixation system presented in this paper differs from those described by Studievic et al. (1971) and Vargues and Henley (1974). In the one developed here only one sampler is used with a wheel presenting a double row of cups. This system can assay 60 samples per h and can be modified to double t...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(76)90183-6

    authors: Coursaget P,Maupas P,Goudeau A,Millman I

    更新日期:1976-01-01 00:00:00

  • Use of the periodate oxidation coupling method for the detection of antibody and antibody-producing cells specific for staphylococcal lipoteichoic acid.

    abstract::The periodate oxidation and chromium chloride coupling methods were compared for their ability to sensitize indicator erythrocytes with staphylococcal lipoteichoic acid (LTA) for the detection of specific antibody. Erythrocytes, sensitized with periodiate-activated lipoteichoic acid, were found to be superior for use ...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(80)90070-8

    authors: Beining PR,Flannery GM,Caldes G,Prescott B,Baker PJ

    更新日期:1980-01-01 00:00:00

  • Hematological response of rabbits to chronic, repetitive, severe bleedings for the production of antisera.

    abstract::A study was undertaken to determine whether large amounts of blood (antiserum) could be rapidly removed from rabbits over a two-month period using a simple suction technique. It was found that half a blood volume could be readily removed each week (over 900 ml/rabbit/54 days) without a single fatality. The rabbits eff...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(78)90081-9

    authors: Nerenberg ST,Zedler P,Prasad R,Biskup NS,Pedersen L

    更新日期:1978-01-01 00:00:00

  • Luminescence immunoassay (LIA): a solid-phase immunoassay monitored by chemiluminescence.

    abstract::A luminescence immunoassay (LIA) has been developed utilizing the chemiluminescent luminol reaction with heme as catalyst. Rabbit antibody against human serum albumin was quantitated in antigen coated plastic tubes using commercially available goat anti-rabbit IgG conjugated to horseradish peroxidase which was the sou...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(79)90048-6

    authors: Olsson T,Brunius G,Carlsson HE,Thore A

    更新日期:1979-01-01 00:00:00

  • A new antigen scanning strategy for monitoring HIV-1 specific T-cell immune responses.

    abstract::Delineation of the immune correlates of protection in natural infection or after vaccination is a mandatory step for vaccine development. Although the most recent techniques allow a sensitive and specific detection of the cellular immune response, a consensus on the best strategy to assess their magnitude and breadth ...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/j.jim.2011.09.005

    authors: Malnati MS,Heltai S,Cosma A,Reitmeir P,Allgayer S,Glashoff RH,Liebrich W,Vardas E,Imami N,Westrop S,Nozza S,Tambussi G,Buttò S,Fanales-Belasio E,Ensoli B,Ensoli F,Tripiciano A,Fortis C,Lusso P,Poli G,Erfle V,Hol

    更新日期:2012-01-31 00:00:00

  • Use of real-time PCR to measure Epstein-Barr virus genomes in whole blood.

    abstract::The measurement of the Epstein-Barr viral load in peripheral blood has been recognised as an important way of monitoring the response to treatment in patients with Epstein-Barr virus (EBV)-related malignancies. In particular, EBV load in transplant recipients can be used as a predictive parameter for Post-transplant L...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/s0022-1759(02)00333-2

    authors: Leung E,Shenton BK,Jackson G,Gould FK,Yap C,Talbot D

    更新日期:2002-12-15 00:00:00

  • A highly sensitive, non-radioactive assay for T cell activation in cattle: applications in screening for antigens recognised by CD4(+) and CD8(+) T cells.

    abstract::We describe a highly sensitive, non-radioactive assay for T cell activation, based on the rapid induction of class II MHC expression by constitutively negative bovine endothelial cells, when cultured in the presence of supernatants derived from activated bovine T cells. We demonstrate the effectiveness of this assay i...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/s0022-1759(00)00187-3

    authors: Ballingall KT,Mwangi DM,MacHugh ND,Taracha EL,Totte P,McKeever1 DJ

    更新日期:2000-05-26 00:00:00

  • The use of denaturing conditions for gel diffusion of insoluble epidermal proteins.

    abstract::Modifications of the classical technique for double diffusion in agar plates are presented. These procedures allow insoluble proteins to be dissolved in a variety of denaturing solvents and antibody production specificity to be monitored by the appearance of precipitin lines in agarose gels as in the conventional Ouch...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(77)90192-2

    authors: Lee LD,Baden HP

    更新日期:1977-01-01 00:00:00

  • High efficiency creation of human monoclonal antibody-producing hybridomas.

    abstract::The native human antibody repertoire holds unexplored potential for the development of novel monoclonal antibody therapeutics. Current techniques that fuse immortal cells and primary B-lymphocytes are sub-optimal for the routine production of hybridomas that secrete human monoclonal antibodies. We have found that a mu...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/j.jim.2004.05.005

    authors: Dessain SK,Adekar SP,Stevens JB,Carpenter KA,Skorski ML,Barnoski BL,Goldsby RA,Weinberg RA

    更新日期:2004-08-01 00:00:00

  • Detection of potassium channel KIR4.1 antibodies in Multiple Sclerosis patients.

    abstract::The presence of KIR4.1 antibodies has been proposed to be a characteristic of Multiple Sclerosis (MS). This could have a significant impact on disease management. However, the validation of the initial findings has failed till date. Conflicting results have been attributed to difficulties in isolating the lower-glycos...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/j.jim.2017.03.008

    authors: Marnetto F,Valentino P,Caldano M,Bertolotto A

    更新日期:2017-06-01 00:00:00

  • A europium fluoroimmunoassay for measuring binding of antigen to class II MHC glycoproteins.

    abstract::A dissociation-enhanced lanthanide fluoroimmunoassay employing europium-streptavidin and time-resolved fluorimetry was developed to measure binding of biotin-labeled peptides to class II MHC proteins. Binding of biotin-peptides as measured by this assay was saturable and inhibited in the presence of unlabeled peptide....

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(93)90124-p

    authors: Tompkins SM,Rota PA,Moore JC,Jensen PE

    更新日期:1993-08-09 00:00:00

  • MicroELISA method for measurement of human serum thymosin alpha 1.

    abstract::A microELISA for the estimation of human serum thymosin alpha 1 is described. In this assay, antibody to thymosin alpha 1 is pre-incubated with the standard or serum at 4 degrees C. Unbound antibody in the liquid-phase then binds with solid-phase thymosin alpha 1. The method is sufficiently sensitive for measuring ser...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(85)90163-2

    authors: Weller FE,Mutchnick MG,Keren DF,Goldstein AL,Naylor PH

    更新日期:1985-06-12 00:00:00

  • Differential binding properties of protein A and protein G for dog immunoglobulins.

    abstract::We studied the binding of dog immunoglobulins G, A, M and E to protein A and protein G. Passive cutaneous anaphylaxis (PCA) testing was used for the measurement of dog IgE and enzyme-linked immunosorbent assays (ELISA) were used for the measurements of dog IgG, IgA and IgM. Protein A from lyophilized cells of Staphylo...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(91)90335-d

    authors: Peng ZK,Simons FE,Becker AB

    更新日期:1991-12-15 00:00:00

  • Liposomes as vaccine carriers. Incorporation of soluble and particulate antigens in giant vesicles.

    abstract::Giant liposomes (mean diameter 5.5 microns) composed of egg phosphatidylcholine or distearoyl phosphatidylcholine, phosphatidyl glycerol, cholesterol and triolein were prepared by a double emulsion technique. They were then mixed with model particulate (killed Bacillus subtilis, and killed Bacille Calmette-Guérin) and...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(93)90368-h

    authors: Antimisiaris SG,Jayasekera P,Gregoriadis G

    更新日期:1993-12-03 00:00:00

  • An improved, ultrasensitive method for the detection of IgM oligoclonal bands in cerebrospinal fluid.

    abstract::A new, 10-fold more sensitive method, based on an improved immunofixation technique, has been devised to detect oligoclonal IgM bands in unconcentrated cerebrospinal fluid (CSF). Using agarose gel electrophoresis, 5 microliters of an unconcentrated sample containing oligoclonal bands was separated and blotted on to a ...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/s0022-1759(97)00023-9

    authors: Rijcken CA,Thompson EJ,Teelken AW

    更新日期:1997-04-25 00:00:00

  • Generation of peptide-specific CD8+ T cells by phytohemagglutinin-stimulated antigen-mRNA-transduced CD4+ T cells.

    abstract::Functional analysis of antigen-specific CD8(+) T cells is important for understanding the immune response in various immunological disorders. To analyze CD8(+) T cell responses to a variety of antigens with no readily defined peptides available, we developed a system using CD4(+) phytohemagglutinin (PHA) blasts transd...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/j.jim.2006.05.009

    authors: Naota H,Miyahara Y,Okumura S,Kuzushima K,Akatsuka Y,Hiasa A,Kitano S,Takahashi T,Yuta A,Majima Y,Shiku H

    更新日期:2006-07-31 00:00:00

  • The use of post-source decay in matrix-assisted laser desorption/ionisation mass spectrometry to delineate T cell determinants.

    abstract::The identification of naturally processed peptides presented by molecules of the major histocompatibility complex (MHC) has progressed significantly over the past decade. The elution of peptides from immunoaffinity purified complexes of MHC class I or class II molecules has provided highly specific biochemical informa...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章,评审

    doi:10.1016/s0022-1759(00)00361-6

    authors: Purcell AW,Gorman JJ

    更新日期:2001-03-01 00:00:00

  • BlockLogo: visualization of peptide and sequence motif conservation.

    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

    authors: Olsen LR,Kudahl UJ,Simon C,Sun J,Schönbach C,Reinherz EL,Zhang GL,Brusic V

    更新日期:2013-12-31 00:00:00

  • Computer-assisted analysis of in vivo cytotoxic T cell responses.

    abstract::An approach has been devised for analysing data generated in the study of in vivo cytotoxic T cell responses. The method calculates the total number of lytic units generated in the peritoneal cavity, or in cultures of the lymph nodes draining the rear footpad, following antigenic stimulation. The assessment of group d...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(83)90387-3

    authors: Crispe IN,Gascoigne NR

    更新日期:1983-11-11 00:00:00