Search results for: Sf9
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#33612723 // To Up
Expression and characterization of anticoagulant activity of salivary protein alALP from Asian tiger mosquito Aedes albopictus.Several bioactive molecules isolated from the saliva of blood-sucking arthropods, such as mosquitoes, have been shown to exhibit potential anticoagulant function. We have previously identified a 30kDa allergen named Aegyptin-like protein (alALP), which is highly homologous to Aegyptin, from the salivary glands of female Aedes albopictus (Asian tiger mosquito). In this study, we identified the conserved functional domain of alALP by using bioinformatic tools, and expressed the His-tagged alALP recombinant protein in sf9 insect cells by generation and transfection of a baculoviral expression plasmid carrying the fulllength cDNA of alALP. We purified this recombinant protein and examined its function on the inhibition of blood coagulation. The results showed that the purified His-alALP prolonged the Activated Partial Thromboplastin Time (APTT), Prothrombin Time (PT) and Thrombin Time (TT) in vitro as well as the Bleeding Time (BT) in vivo, which suggest that alALP could be a novel anticoagulant.
X P Li, D Lin, Y Zhang, S Q Chen, H Q Bai, S N Zhang, W Q Liu, S H Liang
1252 related Products with: Expression and characterization of anticoagulant activity of salivary protein alALP from Asian tiger mosquito Aedes albopictus.2x 100ug24 reactions 5 reactions1x96 well plate 5 G100ul1000 TESTS/0.65ml25ml1050ug1 Set1mg
#33582985 // To Up
Membrane Protein Production and Purification from Escherichia coli and Sf9 Insect Cells.A major obstacle to studying membrane proteins by biophysical techniques is the difficulty in producing sufficient amounts of materials for functional and structural studies. To overexpress the target membrane protein heterologously, especially an eukaryotic protein, a key step is to find the optimal host expression system and perform subsequent expression optimization. In this chapter, we describe protocols for screening membrane protein production using bacterial and insect cells, solubilization screening, large-scale production, and commonly used affinity chromatography purification methods. We discuss general optimization conditions, such as promoters and tags, and describe current techniques that can be used in any laboratory without specialized expensive equipment. Especially for insect cells, GFP fusions are particularly useful for localization and in-gel fluorescence detection of the proteins on SDS-PAGE. We give detailed protocols that can be used to screen the best expression and purification conditions for membrane protein study.
Yixin Liu, Ana Pavić, Joshua T Farley, Carine de Marcos Lousa, Adrian Goldman, Vincent L G Postis
2558 related Products with: Membrane Protein Production and Purification from Escherichia coli and Sf9 Insect Cells.100ul1000 TESTS/0.65ml0.1 mg100 100ml5050 10 10 mg1 mg10
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#33536573 2021/02/03 To Up
A new tool for technical standardization of the Ki67 immunohistochemical assay.Ki67, a nuclear proliferation-related protein, is heavily used in anatomic pathology but has not become a companion diagnostic or a standard-of-care biomarker due to analytic variability in both assay protocols and interpretation. The International Ki67 Working Group in breast cancer has published and has ongoing efforts in the standardization of the interpretation of Ki67, but they have not yet assessed technical issues of assay production representing multiple sources of variation, including antibody clones, antibody formats, staining platforms, and operators. The goal of this work is to address these issues with a new standardization tool. We have developed a cell line microarray system in which mixes of human Karpas 299 or Jurkat cells (Ki67) with Sf9 (Spodoptera frugiperda) (Ki67) cells are present in incremental standardized ratios. To validate the tool, six different antibodies, including both ready-to-use and concentrate formats from six vendors, were used to measure Ki67 proliferation indices using IHC protocols for manual (bench-top) and automated platforms. The assays were performed by three different laboratories at Yale and analyzed using two image analysis software packages, including QuPath and Visiopharm. Results showed statistically significant differences in Ki67 reactivity between each antibody clone. However, subsets of Ki67 assays using three clones performed in three different labs show no significant differences. This work shows the need for analytic standardization of the Ki67 assay and provides a new tool to do so. We show here how a cell line standardization system can be used to normalize the staining variability in proliferation indices between different antibody clones in a triple negative breast cancer cohort. We believe that this cell line standardization array has the potential to improve reproducibility among Ki67 assays and laboratories, which is critical for establishing Ki67 as a standard-of-care assay.
Thazin Nwe Aung, Balazs Acs, Jonathan Warrell, Yalai Bai, Patricia Gaule, Sandra Martinez-Morilla, Ioannis Vathiotis, Saba Shafi, Myrto Moutafi, Mark Gerstein, Benjamin Freiberg, Regan Fulton, David L Rimm
1145 related Products with: A new tool for technical standardization of the Ki67 immunohistochemical assay.100Tests1,000 tests96 Tests100 tests100 assays50 assays100400 assays100 assays100 tests
#33527628 2021/02/01 To Up
Two carboxylesterase genes in Plutella xylostella associated with sex pheromones and plant volatiles degradation.Carboxyl/cholinesterases (CCEs) are thought to play a pivotal role in the degradation of sex pheromones and plant-derived odorants in insects, but their exact biochemistry and physiological functions remain unclear.
Mei-Mei Wang, Gui-Jun Long, Huan Guo, Xuan-Zheng Liu, Hong Wang, Youssef Dewer, Zhao-Qun Li, Kun Liu, Qiu-Liang Zhang, Yun-Feng Ma, Peng He, Ming He
2430 related Products with: Two carboxylesterase genes in Plutella xylostella associated with sex pheromones and plant volatiles degradation.4/120 Packing /sleeve/bo96tests4/120 Packing /sleeve/bo4/120 Packing /sleeve/bo100ug1 Set
#33526755 2021/01/22 To Up
Antiviral Treatment Reveals a Cooperative Pathogenicity of Baculovirus and Iflavirus in , a lepidopteran insect.The beet armyworm, , is a serious insect pest infesting various vegetable crops. Two infectious insect viruses, baculovirus and iflavirus, are known to induce epizootic in populations. Indeed, some laboratory colonies appeared to be covertly infected by these viruses. Diagnostic PCR tests detected two kinds of viruses: multiple nucleopolyhedrosis virus (SeMNPV) and iflaviruses (SeIfV1 and SeIfV2). Viral extract from dead larvae of could infect Sf9 cells and produce occlusion bodies (OBs). Feeding OBs to asymptomatic larvae of caused significant viral disease. Interestingly, both SeIfV1 and SeIfV2 increased their titers at late larval stages. Sterilization of laid eggs with 1% sodium hypochloride significantly reduced SeMNPV titers and increased larval survival rate. Double-stranded RNA (dsRNA) specific to SeIfV1 or SeIfV2 significantly reduced viral titers and increased larval survival rate. To continuously feed dsRNA, a recombinant HT115 expressing SeIfV1-dsRNA was constructed with an L4440 expression vector. Adding this recombinant to the artificial diet significantly reduced the SeIfV1 titer and increased larval survival. These results indicate that laboratory colony collapse of is induced by multiple viral infections. In addition, either suppression of SeMNPV or SeIfV infection significantly increased larval survival, suggesting a cooperative pathogenicity between baculovirus and iflavirus against .
Miltan Chandra Roy, Shabbir Ahmed, Md Mahi Imam Mollah, Yonggyun Kim
2873 related Products with: Antiviral Treatment Reveals a Cooperative Pathogenicity of Baculovirus and Iflavirus in , a lepidopteran insect.1 mL1 mL100.00 ug100 μg25100 μg10 100 μg100 μg100ug Lyophilized96 samples
#33518673 // To Up
Evaluation of Combinatory Effects of Plasmodium Circumsporozoite Protein and Complement Regulatory Protein Expression of Recombinant Baculovirus Vectors.Baculovirus vectors (BVs) are safely able to transduce foreign genes and express them in mammalian cells. However, the transduction activity of BVs is strongly reduced by the attack of serum complement, which is one of the major obstacles in the use of BVs for in vivo gene transfer. One strategy to overcome this problem is the display of complement regulatory proteins (CRPs) on BV virions. We previously developed CD46-decay accelerating factor (DAF)-CD59 triple fusion type BV showing potent complement resistance. We also developed BVs expressing Plasmodium circumsporozoite protein (CSP) to enhance transduction efficacy in hepatic cells. In this study, we investigated the combination of CSP and CRPs in a BV system to evaluate transduction efficacy along with complement resistance. To accomplish the combination of CSP and CRPs, we generated insect Sf9 cells stably expressing CRPs, to which CSP type BV was infected. The BVs collected from these infected cells were confirmed to possess both CSP and CRPs in virions. We demonstrated that CSP-CD46-DAF-CD59 type BV, containing both CSP and CD46-DAF-CD59, showed a significant increase in transduction efficacy in human hepatoma HepG2 cells under intact serum exposure compared with control type BV or CSP type BV, retaining both advantages of CSP and CD46-DAF-CD59. Collectively, these results demonstrated that the utilization of stably expressing Sf9 cells to introduce the protein products of interest, e.g., CRPs into BVs, would be useful strategy to generate BVs with novel functions such as resistance against serum complement attack.
Chiaki Kawabata, Yusuke Kawai, Takahiko Tamura
1098 related Products with: Evaluation of Combinatory Effects of Plasmodium Circumsporozoite Protein and Complement Regulatory Protein Expression of Recombinant Baculovirus Vectors.100ul50 2x 100ug100312011001020500
#33514705 2021/01/29 To Up
Structures of monomeric and dimeric PRC2:EZH1 reveal flexible modules involved in chromatin compaction.Polycomb repressive complex 2 (PRC2) is a histone methyltransferase critical for maintaining gene silencing during eukaryotic development. In mammals, PRC2 activity is regulated in part by the selective incorporation of one of two paralogs of the catalytic subunit, EZH1 or EZH2. Each of these enzymes has specialized biological functions that may be partially explained by differences in the multivalent interactions they mediate with chromatin. Here, we present two cryo-EM structures of PRC2:EZH1, one as a monomer and a second one as a dimer bound to a nucleosome. When bound to nucleosome substrate, the PRC2:EZH1 dimer undergoes a dramatic conformational change. We demonstrate that mutation of a divergent EZH1/2 loop abrogates the nucleosome-binding and methyltransferase activities of PRC2:EZH1. Finally, we show that PRC2:EZH1 dimers are more effective than monomers at promoting chromatin compaction, and the divergent EZH1/2 loop is essential for this function, thereby tying together the methyltransferase, nucleosome-binding, and chromatin-compaction activities of PRC2:EZH1. We speculate that the conformational flexibility and the ability to dimerize enable PRC2 to act on the varied chromatin substrates it encounters in the cell.
Daniel Grau, Yixiao Zhang, Chul-Hwan Lee, Marco Valencia-Sánchez, Jenny Zhang, Miao Wang, Marlene Holder, Vladimir Svetlov, Dongyan Tan, Evgeny Nudler, Danny Reinberg, Thomas Walz, Karim-Jean Armache
2142 related Products with: Structures of monomeric and dimeric PRC2:EZH1 reveal flexible modules involved in chromatin compaction.96 wells100ug2 Pieces/Box100ug 50 UG1 ml1 Set5ug1 Set100
#33511494 2021/01/29 To Up
Expression and kinetic analysis of carboxylesterase LmCesA1 from Locusta migratoria.To investigate the biochemical characterization of the carboxylesterase LmCesA1 from Locusta migratoria.
Fei Yin, Wen Ma, Daqi Li, Xueyao Zhang, Jianqin Zhang
1508 related Products with: Expression and kinetic analysis of carboxylesterase LmCesA1 from Locusta migratoria.50 ug 1 module1 module1,000 tests400/kit100ul1 module480/kit1 mg 15 ml
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