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Search results for: Monoclonal Anti Actin produced in mouse

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#34626726   2021/10/06 To Up

An anti-DR5 antibody-curcumin conjugate for the enhanced clearance of activated hepatic stellate cells.

Anti-death receptor 5 (DR5) antibody is a potential therapeutic agent for liver fibrosis because it exhibits anti-fibrotic effects by inducing the apoptosis of activated hepatic stellate cells (HSCs), which are responsible for hepatic fibrogenesis. However, the clinical applications of anti-DR5 antibodies have been limited by their low agonistic activity against DR5. In this study, an anti-DR5 antibody-curcumin conjugate (DCC) was prepared to investigate its effect on the clearance of activated HSCs. The DCC was synthesized through a coupling reaction between a maleimide-functionalized curcumin derivative and a thiolated anti-DR5 antibody. No significant differences were observed in the uptake behaviors of activated HSCs between the bare anti-DR5 antibodies and DCC. Owing to the antioxidant and anti-inflammatory effects of curcumin, DCC-treated HSCs produced much lower levels of reactive oxygen species and inducible nitric oxide synthase than the bare anti-DR5 antibody-treated HSCs. Additionally, the anti-fibrotic effects of DCC on activated HSCs were more prominent than those of the bare anti-DR5 antibodies, as demonstrated by the immunocytochemical analysis of α-smooth muscle actin. DCC preferentially accumulated in the liver after its systemic administration to mice with liver fibrosis. Thus, DCC may serve as a potential therapeutic agent for treating liver fibrosis.
Van Quy Nguyen, Dong Gil You, Chan Ho Kim, Seunglee Kwon, Wooram Um, Byeong Hoon Oh, Jae Yoon An, Jueun Jeon, Jae Hyung Park

1105 related Products with: An anti-DR5 antibody-curcumin conjugate for the enhanced clearance of activated hepatic stellate cells.

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#32022386   2020/02/05 To Up

IL-22 produced by Th22 cells aggravates atherosclerosis development in ApoE mice by enhancing DC-induced Th17 cell proliferation.

Th22 cells are a novel subset of CD4 T cells that primarily mediate biological effects through IL-22, with both Th22 cells and IL-22 being closely associated with multiple autoimmune and chronic inflammatory diseases. In this study, we investigated whether and how Th22 cells affect atherosclerosis. ApoE mice and age-matched C57BL/6J mice were fed a Western diet for 0, 4, 8 or 12 weeks. The results of dynamic analyses showed that Th22 cells, which secrete the majority of IL-22 among the known CD4 cells, play a major role in atherosclerosis. ApoE mice fed a Western diet for 12 weeks and administered recombinant mouse IL-22 (rIL-22) developed substantially larger plaques in both the aorta and aortic root and higher levels of CD3 T cells, CD68 macrophages, collagen, IL-6, Th17 cells, dendritic cells (DCs) and pSTAT3 but lower smooth muscle cell (SMC) α-actin expression than the control mice. Treatment with a neutralizing anti-IL-22 monoclonal antibody (IL-22 mAb) reversed the above effects. Bone marrow-derived DCs exhibited increased differentiation into mature DCs following rIL-22 and ox-LDL stimulation. IL-17 and pSTAT3 were up-regulated after stimulation with IL-22 and ox-LDL in cells cocultured with CD4 T cells and mature DC supernatant, but this up-regulation was significantly inhibited by IL-6mAb or the cell-permeable STAT3 inhibitor S31-201. Thus, Th22 cell-derived IL-22 aggravates atherosclerosis development through a mechanism that is associated with IL-6/STAT3 activation, DC-induced Th17 cell proliferation and IL-22-stimulated SMC dedifferentiation into a synthetic phenotype.
Lei Shi, Qingwei Ji, Ling Liu, Ying Shi, Zhengde Lu, Jing Ye, Tao Zeng, Yan Xue, Zicong Yang, Yu Liu, Jianyong Lu, Xinshun Huang, Qiuwen Qin, Tianzhu Li, Ying-Zhong Lin

2773 related Products with: IL-22 produced by Th22 cells aggravates atherosclerosis development in ApoE mice by enhancing DC-induced Th17 cell proliferation.

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#27335638   2016/03/30 To Up

Monoclonal antibodies against muscle actin isoforms: epitope identification and analysis of isoform expression by immunoblot and immunostaining in normal and regenerating skeletal muscle.

Higher vertebrates (mammals and birds) express six different highly conserved actin isoforms that can be classified in three subgroups: 1) sarcomeric actins, α-skeletal (α-SKA) and α-cardiac (α-CAA), 2) smooth muscle actins (SMAs), α-SMA and γ-SMA, and 3) cytoplasmic actins (CYAs), β-CYA and γ-CYA. The variations among isoactins, in each subgroup, are due to 3-4 amino acid differences located in their acetylated N-decapeptide sequence. The first monoclonal antibody (mAb) against an actin isoform (α-SMA) was produced and characterized in our laboratory in 1986 (Skalli  et al., 1986) . We have further obtained mAbs against the 5 other isoforms. In this report, we focus on the mAbs anti-α-SKA and anti-α-CAA obtained after immunization of mice with the respective acetylated N-terminal decapeptides using the Repetitive Immunizations at Multiple Sites Strategy (RIMMS). In addition to the identification of their epitope by immunoblotting, we describe the expression of the 2 sarcomeric actins in mature skeletal muscle and during muscle repair after micro-lesions. In particular, we analyze the expression of α-CAA, α-SKA and α-SMA by co-immunostaining in a time course frame during the muscle repair process. Our results indicate that a restricted myocyte population expresses α-CAA and suggest a high capacity of self-regeneration in muscle cells. These antibodies may represent a helpful tool for the follow-up of muscle regeneration and pathological changes.
Christine Chaponnier, Giulio Gabbiani

1787 related Products with: Monoclonal antibodies against muscle actin isoforms: epitope identification and analysis of isoform expression by immunoblot and immunostaining in normal and regenerating skeletal muscle.

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#26680363   2015/12/17 To Up

Characterization of the expression of the pro-metastatic Mena(INV) isoform during breast tumor progression.

Several functionally distinct isoforms of the actin regulatory Mena are produced by alternative splicing during tumor progression. Forced expression of the Mena(INV) isoform drives invasion, intravasation and metastasis. However, the abundance and distribution of endogenously expressed Mena(INV) within primary tumors during progression remain unknown, as most studies to date have only assessed relative mRNA levels from dissociated tumor samples. We have developed a Mena(INV) isoform-specific monoclonal antibody and used it to examine Mena(INV) expression patterns in mouse mammary and human breast tumors. Mena(INV) expression increases during tumor progression and to examine the relationship between Mena(INV) expression and markers for epithelial or mesenchymal status, stemness, stromal cell types and hypoxic regions. Further, while Mena(INV) robustly expressed in vascularized areas of the tumor, it is not confined to cells adjacent to blood vessels. Altogether, these data demonstrate the specificity and utility of the anti-Mena(INV)-isoform specific antibody, and provide the first description of endogenous Mena(INV) protein expression in mouse and human tumors.
Madeleine J Oudin, Shannon K Hughes, Nazanin Rohani, Mira N Moufarrej, Joan G Jones, John S Condeelis, Douglas A Lauffenburger, Frank B Gertler

2477 related Products with: Characterization of the expression of the pro-metastatic Mena(INV) isoform during breast tumor progression.

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#25552314   // To Up

A Novel Monoclonal Antibody Against a Synthetic Peptide from β-Actin can React with its Corresponding Protein.

Actin is one of the most widely studied structural and multifunctional housekeeping proteins in eukaryotic cells with important roles in many cell functions. Antibodies against β-actin and other housekeeping gene-encoded proteins are used as internal loading controls in Western blot analyses. The aim of this study was to produce a monoclonal antibody (mAb) against a synthetic peptide derived from N-terminal region of β-actin and to study its reactivity with different organisms. A synthetic peptide, derived from β-actin, was designed and used to produce a mAb by hybridoma technology. The produced antibody (clone 4E5- A10) was purified by an affinity chromatography column followed by characterization of purified mAb using SDS-PAGE, ELISA and Western blot. Our results showed that 4E5-A10 was an IgM and had desired purity and excellent reactivity with the immunizing peptide with an affinity constant of 2.7x10(8) M(-1)>. It could detect a band of about 45 kDa, corresponding to β-actin, in Western blot. Furthermore, it could react in a more sensitive manner and with a wider range of organisms than a known commercial anti β-actin antibody. Our data suggest that 4E5-A10 can act as a sensitive probe for detection of β-actin as an internal loading control, for a wide range of organisms, in Western blot analyses.
Nazila Amini, Ali-Ahmad Bayat, Omid Zarei, Reza Hadavi, Jafar Mahmoudian, Ahmad R Mahmoudi, Maryam Darzi, Hodjattallah Rabbani, Mahmood Jeddi-Tehrani

2062 related Products with: A Novel Monoclonal Antibody Against a Synthetic Peptide from β-Actin can React with its Corresponding Protein.

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#21474822   2011/04/07 To Up

Src phosphorylation of endothelial cell surface intercellular adhesion molecule-1 mediates neutrophil adhesion and contributes to the mechanism of lung inflammation.

The goal of this study was to determine whether tumor necrosis factor α (TNFα)-induced Src activation and intercellular adhesion molecule-1 (ICAM-1) phosphorylation rapidly increase endothelial cell adhesivity and polymorphonuclear leukocyte (PMN) sequestration independently of de novo ICAM-1 synthesis.
Guoquan Liu, Stephen M Vogel, Xiaopei Gao, Kamran Javaid, Guochang Hu, Sergei M Danilov, Asrar B Malik, Richard D Minshall

1446 related Products with: Src phosphorylation of endothelial cell surface intercellular adhesion molecule-1 mediates neutrophil adhesion and contributes to the mechanism of lung inflammation.

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#19228708   2009/02/19 To Up

Sphingosine-1-phosphate and sphingosine kinase are critical for transforming growth factor-beta-stimulated collagen production by cardiac fibroblasts.

Following injury, fibroblasts transform into myofibroblasts and produce extracellular matrix (ECM). Excess production of ECM associated with cardiac fibrosis severely inhibits cardiac function. Sphingosine-1-phosphate (S1P), a bioactive lysophospholipid, regulates the function of numerous cell types. In this study, we determined the role of S1P in promoting pro-fibrotic actions of cardiac fibroblasts (CFs).
Nicole Gellings Lowe, James S Swaney, Kelli M Moreno, Roger A Sabbadini

2176 related Products with: Sphingosine-1-phosphate and sphingosine kinase are critical for transforming growth factor-beta-stimulated collagen production by cardiac fibroblasts.

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#16164753   2005/09/15 To Up

Metallothionein mediates leukocyte chemotaxis.

Metallothionein (MT) is a cysteine-rich, metal-binding protein that can be induced by a variety of agents. Modulation of MT levels has also been shown to alter specific immune functions. We have noticed that the MT genes map close to the chemokines Ccl17 and Cx3cl1. Cysteine motifs that characterize these chemokines are also found in the MT sequence suggesting that MT might also act as a chemotactic factor.
Xiuyun Yin, David A Knecht, Michael A Lynes

1939 related Products with: Metallothionein mediates leukocyte chemotaxis.

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#11180098   // To Up

Production of stromal cell-derived factor 1 by mesothelial cells and effects of this chemokine on peritoneal B lymphocytes.

B1a lymphocytes accumulate and proliferate in the peritoneal cavity. Stromal cell-derived factor 1 (SDF-1) is a chemotactic and growth promoting factor for B cell precursors. It is required for fetal liver B cell lymphopoiesis, which generates mostly B1a lymphocytes. Using immunohistochemistry with an anti-SDF-1 monoclonal antibody, we found that SDF-1 was produced by peritoneal mesothelial cells in adult mice. Peritoneal B1a lymphocytes expressed a functional SDF-1 receptor, as shown by actin polymerization experiments. In vitro, SDF-1 stimulated migration, proliferation of a minority of peritoneal B1a lymphocytes, and prevented apoptosis in a large fraction of cells. B1a cells migrating in response to SDF-1 were largely enriched in the CD5(high)CD43(high)B220(-)CD1d(-) subpopulation. In vivo, neutralization of SDF-1 for 3 weeks significantly decreased the number of peritoneal B1 cells. SDF-1 also acted on peritoneal B2 cells. These findings show that after the cessation of B cell lymphopoiesis in the liver, around birth, the persistence of B1a cells remains SDF-1 dependent, and that SDF-1 production by mesothelial cells plays a role in the peritoneal location of B1a cells. Thus, the role of mesothelial cells for B1a cells in adults may be similar to that of SDF-1-producing biliary ductal plate cells in the fetus, and to that of bone marrow stromal cells for B2 cell precursors.
A Foussat, K Balabanian, A Amara, L Bouchet-Delbos, I Durand-Gasselin, F Baleux, J Couderc, P Galanaud, D Emilie

2021 related Products with: Production of stromal cell-derived factor 1 by mesothelial cells and effects of this chemokine on peritoneal B lymphocytes.

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#10587368   // To Up

Interleukin-8 expression in endometrial stromal cells is regulated by integrin-dependent cell adhesion.

Concentrations of interleukin (IL)-8, a potent chemotactic factor produced by many cell types, are elevated in the peritoneal fluid of women with endometriosis. We investigated whether endometrial stromal cell (ESC) adhesion induces the expression of IL-8 and if this process is integrin-mediated. ESCs were plated onto culture dishes coated with various extracellular matrix (ECM) substrates, such as fibronectin, laminin, collagen IV, and poly-L-lysine, or mouse anti-human integrin beta(1,) and beta(2) monoclonal antibodies. IL-8 expression was induced by adherence of ESCs to fibronectin or collagen IV, but not to poly-L-lysine, a non-integrin-dependent adhesion matrix. Engagement of beta(1)-containing integrins was associated with ESC adhesion and resulted in up-regulation of IL-8 mRNA expression and protein secretion. Disruption of the actin cytoskeleton by treating ESC with cytochalasin D completely blocked the increase of IL-8 that was induced in response to integrin activation. These findings indicate a novel mechanism of IL-8 regulation; cell adhesion to ECM is an important event that leads to stimulation of IL-8 expression, and this process is mediated by integrins.
J A Garcia-Velasco, A Arici

1069 related Products with: Interleukin-8 expression in endometrial stromal cells is regulated by integrin-dependent cell adhesion.

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