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Search results for: Rabbit Anti-APOA1 Polyclonal Antibody, PE-Cy3 Conjugated

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#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 (κ). 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

1978 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

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#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 32 kDa recombinant TSWV-N protein was induced in BL21 (DE3) cells using 1 mM of Isopropyl β-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 9.48 μg/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 1 μg/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 11 hosts tested, higher virus titer was recorded in Nicotiana tabacum (0.270 μg/100 μL), followed by Impatiens balsamiana (0.185 μg/100 μL) and Dahlia pinnata at a low virus tire of 0.083 μg/100 μL. 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 (̴ 0.115 μg) and 0.711 (̴ 0.111 μg) 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. (̴ 0.044-0.069 μg/100 μL); 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

2879 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 Lyophilized100ug Lyophilized1 mg100ug Lyophilized20200ul100ug Lyophilized

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#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 µg/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

2159 related Products with: Rapid Differential Detection of Abrin Isoforms by an Acetonitrile- and Ultrasound-Assisted On-Bead Trypsin Digestion Coupled with LC-MS/MS Analysis.

1 module100ug Lyophilized25ml 50 mg2.5 mg200ml1 module100 50 ug 25 ml1 module

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#33968077   2021/04/23 To Up

A Double-Antibody Sandwich ELISA for Sensitive and Specific Detection of Swine Fibrinogen-Like Protein 1.

Fibrinogen-like protein 1 (FGL1), a member of the fibrinogen family, is a specific hepatocyte mitogen. Recently, it has been reported that FGL1 is the main inhibitory ligand of lymphocyte activating gene 3 (LAG3). Furthermore, the FGL1-LAG3 pathway has a synergistic effect with programmed death 1 (PD-1)/programmed death ligand 1 (PD-L1) pathway and is regarded as a promising immunotherapeutic target. However, swine FGL1 (sFGL1) has not been characterized and its detection method is lacking. In the study, the sFGL1 gene was amplified from the liver tissue of swine and then inserted into a prokaryotic expression vector, pQE-30. The recombinant plasmid pQE30-sFGL1 was transformed into JM109 competent cells. The recombinant sFGL1 was induced expression by isopropyl-β-d-thiogalactoside (IPTG) and the purified sFGL1 was used as an antigen to produce mouse monoclonal antibody (mAb) and rabbit polyclonal antibody (pAb). After identification, a double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) for sensitive and specific detection of sFGL1 was developed. Swine FGL1 in samples was captured by anti-sFGL1 mAb followed by detection with anti-sFGL1 rabbit pAb and HRP-conjugated goat anti-rabbit IgG. The limit of detection of the developed sFLG1-DAS-ELISA is 35 pg/ml with recombinant sFLG1. Besides, it does not show cross-reactivity with the control protein. Then serum samples of PRRSV-negative and -positive pigs were tested with the established DAS-ELISA and calculated according to the equation of y=0.0735x+0.0737. The results showed that PRRSV infection enhanced the serum FGL1 levels significantly. Our research provides a platform for the research on the functional roles of swine FGL1.
Xin Zhang, Haipeng Zhu, Xu Zheng, Yunjie Jiao, Lulu Ning, En-Min Zhou, Yang Mu

2416 related Products with: A Double-Antibody Sandwich ELISA for Sensitive and Specific Detection of Swine Fibrinogen-Like Protein 1.

1 kit0.1 mg25 µg100ug Lyophilized100 ml100 TESTS100ug100μg100ug Lyophilized0.1 ml0.1 mg96 tests

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#33636533   2021/02/20 To Up

Adulteration of cow's milk with buffalo's milk detected by an on-site carbon nanoparticles-based lateral flow immunoassay.

A competitive lateral flow immunoassay using amorphous carbon nanoparticles (CNPs) and non-immunoglobulin antigen has been developed for the rapid detection of adulteration of cow's milk with buffalo's milk. Purified polyclonal antibodies against a specific buffalo's milk protein fraction were conjugated to CNPs and sprayed on a conjugate pad. The test line consisted of buffalo's skimmed milk proteins (1.6 μg/cm), while the control line contained anti-rabbit antibodies raised in goat (0.5 μg/cm). In the test procedure milk sample is mixed with 100 mM borate buffer (pH 8.8 containing 1% BSA and 0.05% Tween 20) and pipetted onto the sample-cum-conjugate pad. A black/grey test line can be observed if the sample is free from buffalo's milk. The sensitivity of the test i.e. no visible test line is 5% adulteration of cow's milk with buffalo's milk. The test has applicability at the milk receiving stations and can be applied to heated milk samples.
Rajan Sharma, Archana Verma, Nitin Shinde, Bimlesh Mann, Kamal Gandhi, Jan H Wichers, Aart van Amerongen

2430 related Products with: Adulteration of cow's milk with buffalo's milk detected by an on-site carbon nanoparticles-based lateral flow immunoassay.

100ug Lyophilized1 kit(96 Wells)100ug Lyophilized100ug Lyophilized100μg100ug Lyophilized100ug100ug Lyophilized100ug Lyophilized100ug Lyophilized100ug Lyophilized

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#33527460   2021/02/01 To Up

Development and evaluation of colloidal gold immunochromatography test strip for rapid diagnosis of nervous necrosis virus in golden grey mullet (Chelon aurata).

A lateral flow immunochromatography strip test, based on antibody-gold nanoparticles specific for nervous necrosis virus (NNV), was developed for rapid, on-site detection of the virus in fish stocks. A monoclonal antibody against NNV was conjugated with colloidal gold as the detector antibody. A rabbit anti-NNV polyclonal antibody and goat anti-mouse IgG antibody were blotted onto the nitrocellulose membrane as the capture antibodies on the test line and control line, respectively. The reaction could be seen by the eye within 15 min and did not cross-react with the other viruses tested. The detection limit of the strip was approximately 10 TCID /ml and had good stability after storage at 4°C for 8 months. When brains of 70 naturally infected golden grey mullet, Chelon aurata, were tested with the strip test, the diagnostic specificity and sensitivity of the test compared to real-time RT-PCR were 100% and 74%, respectively. Therefore, the one-step test strip developed here had high specificity, reproducibility, and stability. This, together with its simplicity to use and rapid detection, without the requirement of sophisticated equipment or specialized skills, makes the strip suitable for pond-side detection of NNV in farmed fish.
Fatemeh Hassantabar, Mohammad J Zorriehzahra, Farid Firouzbakhsh, Kim D Thompson

2879 related Products with: Development and evaluation of colloidal gold immunochromatography test strip for rapid diagnosis of nervous necrosis virus in golden grey mullet (Chelon aurata).

20 ul50 ul100500 tests500 tests20 ul20 ul50 ul20 ul500 tests10050 ul

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#33001312   2020/10/01 To Up

Efficient detection of eukaryotic calcium-sensing receptor (CaSR) by polyclonal antibody against prokaryotic expressed truncated CaSR.

Calcium-sensing receptor (CaSR), which is better known for its action as regulating calcium homeostasis, can bind various ligands. To facilitate research on CaSR and understand the receptor's function further, an in silico designed truncated protein was developed. The resulting protein folding indicated that 99% of predicted three dimensional (3D) structure residues are located in favored and allowed Ramachandran plots. However, it was found that such protein does not fold properly when expressed in prokaryotic host cells. Thioredoxin (Trx) tag was conjugated to increase the final protein's solubility, which could help obtain the soluble antigen with better immunogenic properties. The truncated recombinant proteins were expressed and purified in two forms (Trx-CaSR: RR19 and CaSR: RRJ19). The polyclonal antibody was induced by the rabbit immunization with the form of RR19. Western blot on mouse kidney lysates evidenced the proper immune recognition of the receptor by the produced antibody. The specificity and sensitivity of antibodies were also assayed by immunohistofluorescence. These experiments affirmed antibody's ability to indicate the receptor on the cell surface in native form and the possibility of applying such antibodies in further cellular and tissue assays.
Aghdas Ramezani, Mohammad Javad Rasaee, Amirmohsen Jalaeefar, Ali Hatef Salmanian

1120 related Products with: Efficient detection of eukaryotic calcium-sensing receptor (CaSR) by polyclonal antibody against prokaryotic expressed truncated CaSR.

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