Home > Rabbit Anti-EXPB-2 Polyclonal Antibody, PE-Cy3 Conjugated
Search results for: Rabbit Anti-EXPB-2 Polyclonal Antibody, PE-Cy3 Conjugated
#35142129 // To Up
[Development of a double-antibody sandwich ELISA targeting the receptor binding domain of TcdB toxin of ST11 type of porcine origin].is an important zoonotic intestinal pathogen, which is widely present in humans and a variety of animals. The ST11 type is one of the most widespread and harmful subtypes in the world. As a large country in pig farming, China lacks efficient methods for detecting of porcine origin, leaving hidden dangers for the prevention and control of . The aim of this study was to develop a specific and sensitive double-antibody sandwich ELISA for the epidemiological investigation of ST11 type of porcine origin. Firstly, a 97 kDa receptor binding domain (RBD) was expressed in a prokaryotic host and purified. A hybridoma cell line AE2D3 capable of stably secreting monoclonal antibody targeting the RBD was screened, and the antibody subtype was determined to be IgG2b (Îº). Secondly, a double antibody sandwich ELISA method was developed, where the monoclonal antibody targeting the RBD was used as a detection antibody, and the rabbit polyclonal antibody was used as a capture antibody. The chessboard method was used to determine the matching concentration of the capture antibody and the detection antibody, the antigen coating conditions, the blocking conditions, the incubation conditions for detection antibody and samples to be tested, as well as the reaction conditions of HRP-conjugated and reaction conditions of TMB chromogenic solution. The negative cutoff was 0.152, and no cross-reaction with 13 strains of non-ST11 type was found. The minimum detection concentration of RBD was 8.83 ng/mL. This specific and sensitive double-antibody sandwich ELISA provides a reliable serological detection method for epidemiological investigation of the ST11 type in pig industry.
Wei Liang, Keji Quan, Qin Zhao, Yaomin Wu, Yu Mu, Sanjie Cao
2135 related Products with: [Development of a double-antibody sandwich ELISA targeting the receptor binding domain of TcdB toxin of ST11 type of porcine origin].50μl96tests96T 5 G100ug Lyophilized
#34986448 2021/12/01 To Up
Research Note: Development of a sandwich ELISA for determining plasma growth hormone concentrations in goose.Growth hormone (GH) is required for normal postnatal development in poultry; however, no immunoassay exists to assess its levels in geese plasma, hindering the study of endocrine regulation in this species. We developed a sandwich ELISA to determine the GH concentrations in the plasma of geese. Recombinant goose GH was produced using a eukaryotic expression system and purified for use as the reference standard in ELISA and the antigen for producing the polyclonal antibodies in rabbits. Rabbit anti-goose GH polyclonal antibody was used to coat the wells of the ELISA plate, and its biotinylated form served as the detection antibody. An avidin-conjugated horseradish peroxidase was used to bind the detection antibody and catalyze the chromogenic reaction of 3,3,5,5-tetramethylbenzidine and HO. A sigmoidal curve was fitted to the optical density and the log of the standard GH concentration using the four-parameter logistic model. The sensitivity of the assay was less than 0.156 ng/mL. The intra- and interassay coefficients of variation were less than 9 and 13%, respectively. The response curve of the serially diluted plasma samples from geese exhibited a good parallel relationship with that observed for the reference standards. The assay effectively detected differences in GH concentrations in plasma samples from geese at various physiological stages; thus, it will be useful for future study of their growth and metabolism.
R Chen, R H Guo, M M Lei, H X Zhu, L Y Yan, Z D Shi
1345 related Products with: Research Note: Development of a sandwich ELISA for determining plasma growth hormone concentrations in goose.96T96 assays96T1 kit96 tests0.1ml (1mg/ml)96T1 kit(96 Wells)3x961 mg96T96 Tests/kit
#34896121 2021/12/08 To Up
A sequel study on the occurrence of Tomato spotted wilt virus (TSWV) in cut-chrysanthemum by DAS-ELISA using recombinant nucleocapsid protein to produce polyclonal antiserum.The tomato spotted wilt virus (TSWV) belonging to the genus Orthotospovirus, family Tospoviridae, causes severe necrotic disease in field crops and horticultural crops, resulting in considerable yield loss worldwide. The development of protein-based diagnostics is essential to track the virus transmission and prevent its spread in vegetatively propagated crops such as ornamentals. In this study, nucleocapsid (N) gene of TSWV was cloned in pET 28 a (+) expression vector. Expression of the 32âkDa recombinant TSWV-N protein was induced in BL21 (DE3) cells using 1âmM of Isopropyl Î²-d-1-thiogalactopyranoside (IPTG), and was confirmed through SDS-PAGE and Western blot by fluorescent-labeled secondary antibody. The bacterial cells expressed recombinant TSWV-N protein up to a concentration of 9.48âÎ¼g/mL. The purified protein was used for immunization of a rabbit to produce specific polyclonal antiserum. The TSWV antiserum was conjugated with the enzyme alkaline phosphatase (ALP). Double Antibody Sandwich-Enzyme Linked Immunosorbent Assay (DAS-ELISA) was developed and validated against TSWV infected hosts. This antiserum specifically reacted with recombinant N protein as well as TSWV infected hosts, but not with groundnut bud necrosis orthotospovirus (GBNV) as well as capsicum chlorosis orthotospovirus (CaCV) infecting tomato and chilli plants. The coating antibody at 1âÎ¼g/mL concentration and 1:500 dilution of enzyme conjugate were found to be effective and economical in the detection of recombinant N protein of TSWV and the virus present naturally in the infected hosts. Using standardized DAS-ELISA protocol, the TSWV titer also was quantified in artificially inoculated assay hosts. Among 11âhosts tested, higher virus titer was recorded in Nicotiana tabacum (0.270âÎ¼g/100âÎ¼L), followed by Impatiens balsamiana (0.185âÎ¼g/100âÎ¼L) and Dahlia pinnata at a low virus tire of 0.083âÎ¼g/100âÎ¼L. The diagnostic reagents and protocol (DAS-ELISA) are further validated by detecting the infection of TSWV in chrysanthemum stem cuttings from six different nurseries in the hill stations of Tamil Nadu, India. The DAS-ELISA assay experimented on six varieties from four different nurseries revealed that the Mum Yellow variety had a higher percentage of TSWV infection (36 %), which was followed by the Mum White variety (33 %); both collected from Kotagiri Nursery. The same variety exhibited a higher virus titer by DAS-ELISA, an A value range of 0.733 (Ì´ 0.115âÎ¼g) and 0.711 (Ì´ 0.111âÎ¼g) respectively, and a total of 27 % of TSWV infection was confirmed by screening 800 stem cuttings by DAS-ELISA. The presence of TSWV was also detected in 54 (6.75 %) asymptomatic stem cuttings from different locations, and the A value ranged from 0.325 to 0.468. (Ì´ 0.044-0.069âÎ¼g/100âÎ¼L); this is the first reported development of immune-based diagnostics for TSWV in India. This protocol and diagnostics will be highly useful for quarantine purposes while trading large quantities of planting materials.
Senthilraja Chinnaiah, Malathi Varagur Ganesan, Nakkeeran Sevugapperumal, Suganthy Mariappan, Sivakumar Uthandi, Renukadevi Perumal
1458 related Products with: A sequel study on the occurrence of Tomato spotted wilt virus (TSWV) in cut-chrysanthemum by DAS-ELISA using recombinant nucleocapsid protein to produce polyclonal antiserum.100 100ug Lyophilized1 mg100ug Lyophilized100ug Lyophilized1 mg100ug Lyophilized20100ug Lyophilized200ul100ug Lyophilized
#34069935 2021/05/18 To Up
Rapid Differential Detection of Abrin Isoforms by an Acetonitrile- and Ultrasound-Assisted On-Bead Trypsin Digestion Coupled with LC-MS/MS Analysis.The high toxic abrin from the plant is a type II ribosome-inactivating protein toxin with a human lethal dose of 0.1-1.0 Âµg/kg body weight. Due to its high toxicity and the potential misuse as a biothreat agent, it is of great importance to developing fast and reliable methods for the identification and quantification of abrin in complex matrices. Here, we report rapid and efficient acetonitrile (ACN)- and ultrasound-assisted on-bead trypsin digestion method combined with HPLC-MS/MS for the quantification of abrin isoforms in complex matrices. Specific peptides of abrin isoforms were generated by direct ACN-assisted trypsin digestion and analyzed by HPLC-HRMS. Combined with in silico digestion and BLASTp database search, fifteen marker peptides were selected for differential detection of abrin isoforms. The abrin in milk and plasma was enriched by immunomagnetic beads prepared by biotinylated anti-abrin polyclonal antibodies conjugated to streptavidin magnetic beads. The ultrasound-assisted on-bead trypsin digestion method was carried out under the condition of 10% ACN as denaturant solvent, the entire digestion time was further shortened from 90 min to 30 min. The four peptides of T3A, T12A, T15A and T9A were chosen as quantification for total abrin, abrin-a, abrin-b, and abrin-c/d, respectively. The absolute quantification of abrin and its isoforms was accomplished by isotope dilution with labeled AQUA peptides and analyzed by HPLC-MS/MS (MRM). The developed method was fully validated in milk and plasma matrices with quantification limits in the range of 1.0-9.4 ng/mL for the isoforms of abrin. Furthermore, the developed approach was applied for the characterization of abrin isoforms from various fractions from gel filtration separation of the seeds, and measurement of abrin in the samples of biotoxin exercises organized by the Organization for the Prohibition of Chemical Weapons (OPCW). This study provided a recommended method for the differential identification of abrin isoforms, which are easily applied in international laboratories to improve the capabilities for the analysis of biotoxin samples.
Long-Hui Liang, Yang Yang, Shu Geng, Xi Cheng, Hui-Lan Yu, Chang-Cai Liu, Shi-Lei Liu