Abstract：
:Wnt5a is a noncanonical member of the Wnt family of signaling molecules that has been linked to various physiological and pathological processes including cell differentiation, cell migration, cell growth, vascular remodeling, cancer and chronic inflammation. To understand the role of Wnt5a in these processes, it is necessary to determine the function and expression level of Wnt5a. In this study we developed a sensitive and specific sandwich enzyme-linked immunosorbent assay (ELISA) for detecting Wnt5a. We found that a rabbit anti-human Wnt5a is a suitable capture antibody for establishing a sandwich ELISA. We used two systems to detect Wnt5a: (1) goat anti-mouse Wnt5a and horseradish peroxidase (HRP) conjugated F(ab')(2) donkey anti-goat IgG as detection and enzyme-linked antibodies respectively, or (2) biotinylated goat anti-mouse Wnt5a and HRP-streptavidin as detection antibody and enzyme-linked avidin respectively. A sandwich ELISA using either of these systems failed to detect recombinant mouse (rm)-Wnt5a diluted in Hank's balanced salt solution supplemented with Ca(2+) and Mg(2+) and 1% bovine serum albumin (HBBS+, 1% BSA). Addition of polyethylene glycol (PEG) to the HBBS+, buffer during the binding stage of rm-Wnt5a, afforded the detection of rm-Wnt5a. The use of PEG during both the binding of rm-Wnt5a and detection antibody stages of the assay yielded the maximum signal for rm-Wnt5a. The relationship between the ELISA signal and concentration of Wnt5a was linear with an R(2) of 0.9934. In summary, we have developed a specific and sensitive sandwich ELISA that detects rm-Wnt5a.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Kummitha CM,Mayle KM,Christman MA 2nd,Deosarkar SP,Schwartz AL,McCall KD,Kohn LD,Malgor R,Goetz DJdoi
10.1016/j.jim.2009.11.005subject
Has Abstractpub_date
2010-01-31 00:00:00pages
38-44issue
1-2eissn
0022-1759issn
1872-7905pii
S0022-1759(09)00341-Xjournal_volume
352pub_type
杂志文章abstract：:A sensitive in situ hybridization procedure using both digoxigenin and 35S-labeled riboprobes is described that allows detection of single T cells expressing cytokine mRNA species in both single and double label formats. Modifications to existing procedures have been developed that allow in situ hybridization to be pe...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(95)00027-8
更新日期：1995-05-11 00:00:00
abstract：:The human beta2-microglobulin antigen-antibody system was used as a model to illustrate the versatility of a microradioimmunoassay technique using protein A-bearing Staphylococcus aureus Cowan I strain (SACI) bacteria as a non-specific immunoadsorbent in place of a second antibody. Experimental conditions are describe...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(79)90113-3
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(97)00112-9
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(82)90086-2
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2004.08.017
更新日期：2004-11-01 00:00:00
abstract：:The differences between murine and human neutrophils mean that findings in mice may not translate to humans, and therefore an in vivo model with human neutrophils would be an important methodological advance. We generated humanised mice by injecting human cord blood derived CD34+ stem cells into irradiated NOD-scid-γc...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2012.08.005
更新日期：2012-11-30 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2016.06.003
更新日期：2016-09-01 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(92)90033-p
更新日期：1992-12-08 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2006.01.003
更新日期：2006-03-20 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(89)90190-7
更新日期：1989-11-13 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2012.05.022
更新日期：2012-08-31 00:00:00
abstract：:Two commercially available bioreactor systems, CELLine and miniPERM, were evaluated for their ability to support the production of monoclonal antibody (mAb) from a variety of murine hybridoma cell lines. Production and purity of mAbs were compared between the two systems and with mouse ascites tumour fluid generation....
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(02)00081-9
更新日期：2002-06-01 00:00:00
abstract：:Dendritic Cell (DC)-based vaccination approaches in man require a reproducible DC generation method that can be performed in conformity with GMP (Good Manufacturing Practice) guidelines and that circumvents the need for multiple blood drawings to generate DC. To this end we modified our previously described method to ...
journal_title：Journal of immunological methods
pub_type： 杂志文章,评审
doi：10.1016/s0022-1759(98)00208-7
更新日期：1999-02-01 00:00:00
abstract：:A new method for measuring the uptake of Staphylococcus aureus by human polymorphonuclear neutrophils (PMN) using flow cytometry (FCM) is described. Bacteria were labeled with fluorescein isothiocyanate (FITC) and incubated with PMN in a suspension assay. At the end of the assay, phagocytosis was arrested by the addit...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(89)90161-0
更新日期：1989-07-26 00:00:00
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journal_title：Journal of immunological methods
pub_type： 信件
doi：10.1016/j.jim.2014.07.011
更新日期：2014-12-01 00:00:00
abstract：:Complement activation is a key component of the pathogenesis of immune-mediated tissue damage in many diseases. Assessment of complement activation in current practice is largely based on the measurement of intact C3 and C4 or the determination of complement haemolytic function. These parameters reflect activation onl...
journal_title：Journal of immunological methods
pub_type： 杂志文章,评审
doi：10.1016/0022-1759(93)90063-d
更新日期：1993-01-04 00:00:00
abstract：:We describe an intracellular cytokine staining, flow cytometry based technique for quantitative and qualitative assessment of CD8+ T lymphocyte-mediated antigen specific immune responses. This adaptation of the technique first described by Suni et al. [J. Immunol. Methods, 212 (1998) 89; Cytometry, 40 (2000) 60] utili...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(02)00025-x
更新日期：2002-04-01 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2013.01.004
更新日期：2013-03-29 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2016.02.001
更新日期：2016-04-01 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(95)00032-6
更新日期：1995-06-14 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(89)90179-8
更新日期：1989-11-13 00:00:00
abstract：:Near-infrared Raman spectroscopy may provide a highly sensitive, noninvasive means to identify activation status of leukocytes. The purpose of the current study was to establish Raman spectroscopic characteristics of T cell activation. Activation of the RsL.11 T cell clone in vitro with Con A resulted in specific decr...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2004.11.020
更新日期：2005-02-01 00:00:00
abstract：:In the course of studying the susceptibility of mice to Salmonella infection, we discovered that C3H/HeJ mice differ from other strains in both their susceptibility to infection and their response to prior vaccination. Others have noted the absence of the usual B cell stimulation by LPS in these mice. To investigate t...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(77)90108-9
更新日期：1977-01-01 00:00:00
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
更新日期：1980-01-01 00:00:00
abstract：:Monoclonal antibodies have been prepared against a synthetic peptide with a sequence corresponding to a repeated hydrophilic region of the protein core of the human MUC-2 gastrointestinal mucin. Peptide conjugates, prepared by glutaraldehyde cross-linking with keyhole limpet haemocyanin (KLH) and bovine serum albumin ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(93)90168-7
更新日期：1993-02-26 00:00:00
abstract：:Monoclonal antibodies (Mab) are the fastest growing group of biopharmaceuticals in development. For production in mammalian cells, the four polypeptide chains of the immunoglobulin diheterotetramer must be assembled prior to exit from the endoplasmic reticulum. Various recombinant Mab expression vectors have been deve...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2006.10.010
更新日期：2007-01-10 00:00:00
abstract：:2-Methoxyestradiol (2ME), an end-metabolite of 17beta-estradiol, is an antiproliferative agent that is currently being tested in clinical trials for cancer treatment. We hereby report that sub-cytotoxic concentrations of 2ME influence the in vitro proliferation of human peripheral blood B lymphocytes. More surprisingl...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2010.02.012
更新日期：2010-04-15 00:00:00
abstract：:Synovial fluid (SF) is a difficult biological matrix to analyse due to its complex non-Newtonian nature. This can result in poor assay repeatability and potentially inefficient use of precious samples. This study assessed the impact of SF treatment by hyaluronidase and/or dilution on intra-assay precision using the Lu...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2012.08.012
更新日期：2012-12-14 00:00:00
abstract：:Conditions for the reproducible measurement of thymic hormonal effect on the functional activity of human peripheral blood lymphocytes (PBL) were determined using a mixed lymphocyte-tumor culture assay. Three thymic hormonal preparations (thymopoietin, thymic humoral factor and TP-1) were tested by this assay and foun...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(80)90312-9
更新日期：1980-01-01 00:00:00
abstract：:Enzyme immunoassay techniques are widely use to quantify various antigens and antibodies. The final step of these techniques (i.e. enzyme reaction) may be carried out in several ways (e.g. chromogenic, fluorogenic, or radioactive substrate and thermometric measurement). This paper compares the effectiveness of the chr...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(82)90188-0
更新日期：1982-01-01 00:00:00
