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Validation of a monoclonal antibody directed against the human sphingosine 1-phosphate receptor type 1
The sphingosine 1-phosphate receptor kind 1 (S1PR1) has a number of vital features, together with stabilizing endothelial barrier and sustaining lymphocyte circulation. These features are critically depending on the regulation of S1PR1 cell floor expression. Presently obtainable antibodies in opposition to human S1PR1 are usually not in a position to decide up cell floor expression on residing cells by movement cytometry as a consequence of intracellular epitopes or unspecific binding.
Right here we describe the era of a mouse monoclonal antibody particular for the N-terminal area of human S1PR1. It has an immunoglobulin M (IgM) kappa isotype and detects cell floor expression of recombinant human S1PR1 on overexpressing cells. As a consequence of unspecific intracellular cell staining, it can’t be used for staining of useless cells and tissue slides or in microscopic analyses.
It’s also not appropriate for Western blot evaluation and immunoprecipitation. Nonetheless, the antibody can stain for endogenous S1PR1 on human endothelial cell traces and first human umbilical vein endothelial cells (HUVEC). Incubation of those cells with numerous S1PR1 agonists revealed potent S1PR1 internalization, which was not the case with the particular antagonist W146.
Surprisingly, human T and B cells remoted from blood and palatine tonsils didn’t present particular staining, demonstrating considerably decrease endogenous S1PR1 floor expression on lymphocytes than on endothelial cells.
Sphingosine-1-phosphate receptor modulator FTY720 attenuates experimental myeloperoxidase-ANCA vasculitis in a T cell-dependent method
Sphingosine-1-phosphate (S1P) is a pleiotropic lysosphingolipid derived from the metabolism of plasma membrane lipids. The interplay between S1P and its ubiquitously expressed G-protein-coupled receptors (S1PR1-5) is essential in lots of pathophysiological processes. Rising proof recommended a possible position for S1P receptors in anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV).
Within the current research, we investigated the consequences of three totally different S1P receptors modulators in a acknowledged rat mannequin of experimental autoimmune vasculitis (EAV). The consequences of therapies had been evaluated with clinico-pathological parameters together with hematuria, proteinuria, crescent formation, pulmonary hemorrhage, and so on.
In vitro practical research had been carried out in a Jurkat T-cell line following stimulations of serum from myeloperoxidase-AAV sufferers. We discovered that solely the FTY720 remedy considerably alleviated hematuria and proteinuria, and diminished glomerular crescent formation, renal tubulointerstitial lesions and pulmonary hemorrhage in EAV.
The attenuation was accompanied by much less renal T-cell infiltration, up-regulated mRNA of S1PR1 and down-regulated IL-1β in kidneys, however not altered circulating ANCA ranges, suggesting that the therapeutic results of FTY720 had been B-cell impartial. Additional in vitro research demonstrated that FTY720 incubation might considerably inhibit the proliferation, adhesion, and migration, and enhance apoptosis of T cells.
In conclusion, the S1P modulator FTY720 might attenuate EAV by means of the discount and inhibition of T cells, which could develop into a novel remedy of ANCA-associated vasculitis.
Inflammatory Circumstances Disrupt Constitutive Endothelial Cell Barrier Stabilization by Assuaging Autonomous Secretion of Sphingosine 1-Phosphate.
The breakdown of the endothelial cell (EC) barrier contributes considerably to sepsis mortality. Sphingosine 1-phosphate (S1P) is without doubt one of the simplest EC barrier-stabilizing signaling molecules. Stabilization is principally transduced by way of the S1P receptor kind 1 (S1PR1). Right here, we exhibit that S1P was autonomously produced by ECs. S1P secretion was considerably increased in main human umbilical vein endothelial cells (HUVEC) in comparison with the endothelial cell line EA.hy926.
Constitutive barrier stability of HUVEC, however not EA.hy926, was considerably compromised by the S1PR1 antagonist W146 and by the anti-S1P antibody Sphingomab. HUVEC and EA.hy926 differed within the expression of the S1P-transporter Spns2, which allowed HUVEC, however not EA.hy926, to secrete S1P into the extracellular house. Spns2 poor mice confirmed elevated serum albumin leakage in bronchoalveolar lavage fluid (BALF).
Lung ECs remoted from Spns2 poor mice revealed elevated leakage of fluorescein isothiocyanate (FITC) labeled dextran and decreased resistance in electrical cell-substrate impedance sensing (ECIS) measurements. Spns2 was down-regulated in HUVEC after stimulation with pro-inflammatory cytokines and lipopolysaccharides (LPS), which contributed to destabilization of the EC barrier.
Our work suggests a brand new mechanism for barrier integrity upkeep. Secretion of S1P by EC by way of Spns2 contributed to constitutive EC barrier upkeep, which was disrupted below inflammatory circumstances by way of the down-regulation of the S1P-transporter Spns2.
The mechanism of the premetastatic area of interest facilitating colorectal most cancers liver metastasis generated from myeloid-derived suppressor cells induced by the S1PR1-STAT3 signaling pathway.
The tumor-derived elements concerned within the growth and accumulation of myeloid-derived suppressor cells (MDSCs) in metastatic dissemination of colorectal most cancers (CRC) to the liver has not been studied. Immunohistochemistry was used to detect sphingosine-1-phosphate receptor 1 (S1PR1) and sign transducer and activator of transcription-3 (STAT3) in human colorectal tumors. IL-6 and interferon-γ had been detected by enzyme-linked immunosorbent assay (ELISA).
Tumor progress, invasion, and migration had been evaluated by MTT, transwell, and wound therapeutic assays, respectively. Subcutaneous tumor-bearing and CRC liver metastasis (CRLM) nude mouse fashions had been constructed. The proportion of MDSCs was measured utilizing multicolor movement cytometry.
Western blot assay was used to guage S1PR1 and p-STAT3 expression in MDSCs after separation from the liver and tumor by magnetic antibody. T-cell suppression assay was detected by carboxyfluorescein succinimidyl ester (CFSE). Aberrant co-expressed S1PR1 and p-STAT3 was correlated with metachronous liver metastasis and poor prognosis in CRC. A mutual activation loop between S1PR1 and STAT3 can improve CRC cell proliferation, migration, and invasion in vitro and in vivo.
The expression of p-STAT3 and its downstream proteins might be regulated by S1PR1. p-STAT3 was the dependent signaling pathway of S1PR1 within the promotion of cell progress and liver metastasis in CRC. The extent of IL-6 and the related MDSCs stimulated by the S1PR1-STAT3 correlated with the variety of liver metastatic nodes within the CRLM mouse fashions and sufferers. Elevated CD14+HLA-DR-/low MDSCs from CRLM sufferers inhibited autologous T-cell proliferation and predict poor prognosis.
The S1PR1-STAT3-IL-6-MDSCs axis operates in each tumor cells and MDSCs concerned within the promotion of progress and liver metastasis in CRC. MDSCs induced by S1PR1-STAT3 in CRC cells fashioned the premetastatic area of interest within the liver can promote organ-specific metastasis.
Sphingosine 1-phosphate however not Fingolimod protects neurons in opposition to excitotoxic cell dying by inducing neurotrophic gene expression in astrocytes.
Sphingosine 1-phosphate (S1P) is a necessary lipid metabolite that indicators by means of a household of 5 G-protein coupled receptors, S1PR1-S1PR5, to control cell physiology. The a number of sclerosis drug Fingolimod is a potent S1P receptor agonist that causes peripheral lymphopenia. Latest analysis has demonstrated direct neuroprotective properties of FTY720 in a number of neurodegenerative paradigms, nonetheless neuroprotective properties of the native ligand S1P haven’t been established.
We aimed to determine the importance of neurotrophic issue up-regulation by S1P for neuroprotection, evaluating S1P with FTY720. S1P induced brain-derived neurotrophic issue, leukemia inhibitory issue, platelet-derived progress issue B and heparin-binding EGF-like progress issue gene expression in main human and murine astrocytes, however not in neurons, and to a a lot larger extent than FTY720.
Accordingly, S1P however not FTY720 protected cultured neurons in opposition to excitotoxic cell dying in a main murine neuron-glia co-culture mannequin, and a neutralizing antibody to LIF blocked this S1P-mediated neuroprotection. Antagonists of S1PR1 and S1PR2 each inhibited S1P-mediated neurotrophic gene induction in human astrocytes, indicating that simultaneous activation of each receptors is required.
S1PR1 Antibody |
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| 1-CSB-PA193689 | Cusabio |
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Description: A polyclonal antibody against S1PR1. Recognizes S1PR1 from Human, Mouse, Rat. This antibody is Unconjugated. Tested in the following application: ELISA, WB;ELISA:1:2000-1:5000, WB:1:500-1:2000 |
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S1PR1 Antibody |
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| 1-CSB-PA241754 | Cusabio |
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Description: A polyclonal antibody against S1PR1. Recognizes S1PR1 from Human, Mouse, Rat. This antibody is Unconjugated. Tested in the following application: ELISA, WB;ELISA:1:2000-1:5000, WB:1:500-1:2000 |
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S1PR1 Antibody |
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| 1-CSB-PA007898 | Cusabio |
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Description: A polyclonal antibody against S1PR1. Recognizes S1PR1 from Human. This antibody is Unconjugated. Tested in the following application: WB, IHC, IF, ELISA;WB:1/500-1/2000.IHC:1/100-1/300.IF:1/200-1/1000.ELISA:1/20000 |
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S1PR1 Antibody |
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| 1-CSB-PA020650GA01HU | Cusabio |
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Description: A polyclonal antibody against S1PR1. Recognizes S1PR1 from Human, Mouse, Rat. This antibody is Unconjugated. Tested in the following application: ELISA, WB |
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S1PR1 Antibody |
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| 1-CSB-PA020650LA01HU | Cusabio |
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Description: A polyclonal antibody against S1PR1. Recognizes S1PR1 from Human, Mouse. This antibody is Unconjugated. Tested in the following application: ELISA, WB, IHC, IF; Recommended dilution: WB:1:500-1:2000, IHC:1:20-1:200, IF:1:50-1:200 |
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S1PR1 Antibody |
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| 1-CSB-PA002247 | Cusabio |
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Description: A polyclonal antibody against S1PR1. Recognizes S1PR1 from Human, Mouse, Rat. This antibody is Unconjugated. Tested in the following application: WB, IF, ELISA;WB:1/500-1/2000.IF:1/200-1/1000.ELISA:1/5000 |
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S1pr1 Antibody |
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| F52393-0.4ML | NSJ Bioreagents | 0.4 ml | EUR 322.15 |
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Description: Receptor for the lysosphingolipid sphingosine 1-phosphate (S1P). S1P is a bioactive lysophospholipid that elicits diverse physiological effect on most types of cells and tissues. This inducible epithelial cell G-protein-coupled receptor may be involved in the processes that regulate the differentiation of endothelial cells. Seems to be coupled to the G(i) subclass of heteromeric G proteins. |
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S1pr1 Antibody (Zebrafish) |
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| F52393-0.08ML | NSJ Bioreagents | 0.08 ml | EUR 140.25 |
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Description: Receptor for the lysosphingolipid sphingosine 1-phosphate (S1P). S1P is a bioactive lysophospholipid that elicits diverse physiological effect on most types of cells and tissues. This inducible epithelial cell G-protein-coupled receptor may be involved in the processes that regulate the differentiation of endothelial cells. Seems to be coupled to the G(i) subclass of heteromeric G proteins. |
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S1PR1 Conjugated Antibody |
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| C37288 | SAB | 100ul | EUR 476.4 |
S1PR1 Polyclonal Antibody |
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| A70547 | EpiGentek |
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S1PR1 Polyclonal Antibody |
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| A50464 | EpiGentek |
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S1PR1 Antibody, HRP conjugated |
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| 1-CSB-PA020650LB01HU | Cusabio |
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Description: A polyclonal antibody against S1PR1. Recognizes S1PR1 from Human. This antibody is HRP conjugated. Tested in the following application: ELISA |
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S1PR1 Antibody, FITC conjugated |
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| 1-CSB-PA020650LC01HU | Cusabio |
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Description: A polyclonal antibody against S1PR1. Recognizes S1PR1 from Human. This antibody is FITC conjugated. Tested in the following application: ELISA |
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anti- S1PR1/EDG1 antibody |
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| FNab07568 | FN Test | 100µg | EUR 658.5 |
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Description: Antibody raised against S1PR1/EDG1 |
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S1PR1 Antibody, Biotin conjugated |
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| 1-CSB-PA020650LD01HU | Cusabio |
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Description: A polyclonal antibody against S1PR1. Recognizes S1PR1 from Human. This antibody is Biotin conjugated. Tested in the following application: ELISA |
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Polyclonal S1PR1 / EDG1 / S1P1 Antibody |
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| APR02818G | Leading Biology | 0.05ml | EUR 580.8 |
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Description: A polyclonal antibody raised in Rabbit that recognizes and binds to Human S1PR1 / EDG1 / S1P1 . This antibody is tested and proven to work in the following applications: |
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S1PR1 Polyclonal Antibody, HRP Conjugated |
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| A50465 | EpiGentek |
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S1PR1 Polyclonal Antibody, FITC Conjugated |
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| A50466 | EpiGentek |
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S1PR1 Polyclonal Antibody, Biotin Conjugated |
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| A50467 | EpiGentek |
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Sphingosine 1 Phosphate Receptor 1 (S1PR1) Antibody |
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| 20-abx242381 | Abbexa |
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Sphingosine 1 Phosphate Receptor 1 (S1PR1) Antibody |
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| 20-abx242382 | Abbexa |
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Sphingosine 1 Phosphate Receptor 1 (S1PR1) Antibody |
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| 20-abx174618 | Abbexa |
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Sphingosine 1 Phosphate Receptor 1 (S1PR1) Antibody |
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| 20-abx178450 | Abbexa |
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Sphingosine 1 Phosphate Receptor 1 (S1PR1) Antibody |
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| 20-abx178451 | Abbexa |
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Sphingosine 1 Phosphate Receptor 1 (S1PR1) Antibody |
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| 20-abx178452 | Abbexa |
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Sphingosine 1 Phosphate Receptor 1 (S1PR1) Antibody |
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| 20-abx324887 | Abbexa |
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Sphingosine 1 Phosphate Receptor 1 (S1PR1) Antibody |
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| 20-abx328149 | Abbexa |
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Polyclonal S1PR1 / EDG1 / S1P1 Antibody (aa25-40) |
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| APR02182G | Leading Biology | 0.05mg | EUR 580.8 |
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Description: A polyclonal antibody raised in Rabbit that recognizes and binds to Human S1PR1 / EDG1 / S1P1 (aa25-40). This antibody is tested and proven to work in the following applications: |
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Sphingosine 1-Phosphate Receptor 1 (S1PR1) Antibody |
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| 20-abx318453 | Abbexa |
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Sphingosine 1-Phosphate Receptor 1 (S1PR1) Antibody |
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| abx237568-100ug | Abbexa | 100 ug | EUR 610.8 |
Sphingosine 1-Phosphate Receptor 1 (S1PR1) Antibody |
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| abx218733-100ug | Abbexa | 100 ug | EUR 526.8 |
Sphingosine 1-Phosphate Receptor 1 (S1PR1) Antibody |
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| abx147509-100ug | Abbexa | 100 ug | EUR 526.8 |
Sphingosine 1-phosphate receptor 1 (S1PR1) Antibody |
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| abx415284-01mg | Abbexa | 0.1 mg | EUR 693.6 |
Sphingosine-1-Phosphate Receptor 1 (S1PR1) Antibody |
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| 20-abx115784 | Abbexa |
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Sphingosine 1-Phosphate Receptor 1 (S1PR1) Antibody (HRP) |
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| 20-abx305226 | Abbexa |
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Sphingosine 1-Phosphate Receptor 1 (S1PR1) Antibody (FITC) |
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| 20-abx305227 | Abbexa |
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Sphingosine 1-Phosphate Receptor 1 (S1PR1) Antibody (Biotin) |
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| 20-abx305228 | Abbexa |
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S1PR1 siRNA |
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| 20-abx904763 | Abbexa |
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S1PR1 siRNA |
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| 20-abx932372 | Abbexa |
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S1PR1 siRNA |
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| 20-abx932373 | Abbexa |
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S1PR1-MO-1 |
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| HY-U00366 | MedChemExpress | 5mg | EUR 5415.6 |
S1PR1 Rabbit pAb |
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| A12935-100ul | Abclonal | 100 ul | EUR 369.6 |
S1PR1 Rabbit pAb |
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| A12935-200ul | Abclonal | 200 ul | EUR 550.8 |
S1PR1 Rabbit pAb |
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| A12935-20ul | Abclonal | 20 ul | EUR 219.6 |
S1PR1 Rabbit pAb |
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| A12935-50ul | Abclonal | 50 ul | EUR 267.6 |
S1PR1 cloning plasmid |
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| CSB-CL020650HU-10ug | Cusabio | 10ug | EUR 279.6 |
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Description: A cloning plasmid for the S1PR1 gene. |
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S1PR2 signaling was transduced by means of Gα13 and the small GTPase Rho, and was mandatory for up-regulation and activation of the transcription elements FOS and JUN, which regulate LIF, BDNF and HBEGF transcription. In abstract, we present that S1P protects hippocampal neurons in opposition to excitotoxic cell dying by means of up-regulation of neurotrophic gene expression, significantly LIF, in astrocytes.
