Phospho-S6 (Ser235, Ser236) Monoclonal Antibody (cupk43k), PE-Cyanine7

货号：25-9007-41,25-9007-42

规格：25 Tests,100 Tests

价格：1237,3202

产品类型：流式抗体

品牌：eBioscience

物种：人/小鼠

宿主：小鼠

抗体亚型：IgG1, kappa

克隆号：cupk43k

荧光染料：PE-Cy7

类型:	单抗	同型对照：	Mouse IgG1 kappa Isotype Control (P3.6.2.8.1), PE-Cyanine7
浓度：	5 µL/Test	用法：	5 µL (0.125 µg)/test(Flow)

产品详细信息Description: This cupk43k monoclonal antibody recognizes human and mouse ribosomal protein S6 (also known as 40S ribosomal protein S6, phosphoprotein NP33, RPS6, RS6, S6) when phosphorylated on serine 235 (S235, human) and serine 236 (S236, mouse). Ribosomal protein S6 is a component of the 40S subunit of the ribosome and is phosphorylated at multiple sites following stimulation of cells by growth factors, tumor promoting agents, or mitogens. Phosphorylation of ribosomal protein S6 by p70S6K andPKDCD results in upregulation of the translation of RNA coding for proteins involved in cell cycle entry. Ribosomal protein S6 is dephosphorylated upon growth arrest.The specificity of the cupk43k monoclonal antibody was determined by western blotting.Applications Reported: This cupk43k antibody has been reported for use in intracellular staining followed by flow cytometric analysis.Applications Tested: This cupk43k antibody has been pre-titrated and tested by intracellular staining and flow cytometric analysis of stimulated normal peripheral blood cells. This can be used at 5 µL (0.125 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.Use of Protocol A: Two-step protocol: intracellular (cytoplasmic) proteins allows for the greatest flexibility for detection of surface and intracellular (cytoplasmic) proteins. Use of Protocol B: One-step protocol: intracellular (nuclear) proteins is recommended for staining of transcription factors in conjunction with surface and phosphorylated intracellular (cytoplasmic) proteins. Protocol C: Two-step protocol: Fixation/Methanol allows for the greatest discrimination of phospho-specific signaling between unstimulated and stimulated samples, but with limitations on the ability to stain specific surface proteins (refer to "Clone Performance Following Fixation/Permeabilization" located in the Best Protocols Section under the Resources tab online). All Protocols can be found in the "Staining Intracellular Antigens for Flow Cytometry Protocol" located in the Best Protocols Section under the Resources tab online.Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.Excitation: 488-561 nm; Emission: 775 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.Filtration: 0.2 µm post-manufacturing filtered.靶标信息Ribosomes, the organelles that catalyze protein synthesis, consist of a small 40S subunit and a large 60S subunit. Together these subunits are composed of 4 RNA species and approximately 80 structurally distinct proteins. This gene encodes a cytoplasmic ribosomal protein that is a component of the 40S subunit. The protein belongs to the S6E family of ribosomal proteins. It is the major substrate of protein kinases in the ribosome, with subsets of five C-terminal serine residues phosphorylated bydifferent protein kinases. Phosphorylation is induced by a wide range of stimuli, including growth factors, tumor-promoting agents, and mitogens. Dephosphorylation occurs at growth arrest. The protein may contribute to the control of cell growth and proliferation through the selective translation of particular classes of mRNA. As is typical for genes encoding ribosomal proteins, there are multiple processed pseudogenes of this gene dispersed through the genome.

数据

Phospho-S6 (Ser235, Ser236) Antibody (25-9007-42) in FlowIntracellular staining of freshly-harvested (left), unstimulated (middle), or 30-minute LPS-stimulated (right) normal human peripheral blood cells with Anti-Human CD14 eFluor® 450 (Product # 48-0149-42) and Anti-Human/Mouse phospho-S6 Ribosomal (S235/S236) PE-Cyanine7, using the Intracellular Fixation and Permeabilization Buffer Set (Product # 88-8824-00) and protocol. Cells in the lymphocyte/ monocyte gate were used for analysis.

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参考文献：

1.Cell reportsIFNα Impairs Autophagic Degradation of mtDNA Promoting Autoreactivity of SLE Monocytes in a STING-Dependent Fashion."25-9007 was used in Flow cytometry/Cell sorting to demonstrate IFNα-mediated deregulation of mitochondrial metabolism and impairment of autophagic degradation, leading to cytosolic accumulation of mtDNA that is sensed via stimulator of interferon genes to promote induction of autoinflammatory DCs."AuthorsGkirtzimanaki K,Kabrani E,Nikoleri D,Polyzos A,Blanas A,Sidiropoulos P,Makrigiannakis A,Bertsias G,Boumpas DT,Verginis P

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