Phospho-p38 MAPK (Thr180, Tyr182) Monoclonal Antibody (4NIT4KK), PerCP-eFluor 710

货号：46-9078-41,46-9078-42

规格：25 Tests,100 Tests

价格：1394,3186

产品类型：流式抗体

品牌：eBioscience

物种：人/小鼠

宿主：小鼠

抗体亚型：IgG2b, kappa

克隆号：4NIT4KK

荧光染料：PerCP-eFluor™ 710

类型:	单抗	同型对照：	Mouse IgG2b kappa Isotype Control (eBMG2b), PerCP-eFluor 710
浓度：	5 µL/Test	用法：	5 µL (0.03 µg)/test(Flow)

产品详细信息Description: This 4NIT4KK monoclonal antibody recognizes human and mouse p38 mitogen-activated protein kinase (MAPK) when phosphorylated on T180/Y182. p38 MAPK belongs to a family of conserved serine/threonine protein kinases that are phosphorylated and activated in response to numerous stress stimuli including TLR ligands (such as LPS), osmotic shock, heat shock, UV irradiation, and inflammatory cytokines. There are four p38 MAPK members that include p38 alpha, p38 beta, p38 gamma, and p38 delta. The primary activators of p38 MAPK are MKK3/4 and MKK6. Several downstream targets of p38 MAPK have been identified including MK2/3, p53, ATF-2, and ETS1. p38 MAPK can be negatively regulated by the chemical inhibitor SB203580. Specificity of this 4NIT4KK clone was determined by ELISA, flow cytometry, and western blotting.Applications Reported: This 4NIT4KK antibody has been reported for use in intracellular staining followed by flow cytometric analysis.Applications Tested: This 4NIT4KK antibody has been pre-titrated and tested by intracellular staining followed by flow cytometric analysis of stimulated normal human peripheral blood cells. This can be used at 5 µL (0.03 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.Staining Protocol: All protocols work well for this monoclonal antibody. 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 Flow Cytometry Protocols: "Staining Intracellular Antigens for Flow Cytometry Protocol" located in the Best Protocols Section under the Resources tab online.PerCP-eFluor® 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome.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 nm; Emission: 710 nm; Laser: Blue Laser.Filtration: 0.2 µm post-manufacturing filtered.靶标信息p38 MAPK alpha (MAPK14) and p38 MAPK beta (MAPK11) are serine/threonine kinases which act as an essential component of the MAPkinase signal transduction pathway. P38 MAPK alpha and P38 MAPK beta are two of the four p38 MAPKs which play an important role in the cascades of cellular responses evoked by extracellular stimuli such as proinflammatory cytokines or physical stress leading to direct activation of transcription factors. Accordingly, p38 MAPKs phosphorylate a broad range of proteins and it has been estimated that they may have approximately 200 to 300 substrates each. MAPK11 functions are mostly redundant with those of MAPK14. Some of the targets are downstream kinases which are activated through phosphorylation and further phosphorylate additional targets.

数据

Phospho-p38 MAPK (Thr180, Tyr182) Antibody (46-9078-41) in FlowNormal human peripheral blood cells were untreated (left) or treated for 30 minutes with anisomycin (right) and stained intracellularly with Anti-Human CD11b PE-Cyanine7 (Product # 25-0118-42) and Anti-Human/Mouse phospho-p38 MAPK (T180/Y182) PerCP-eFluor® 710 using the Intracellular Fixation and Permeabilization Buffer set (Product # 88-8824-00) and protocol. Cells in the monocyte gate was used for analysis.

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参考文献：

1.Stem cell reportsEndogenous DNA Damage Leads to p53-Independent Deficits in Replicative Fitness in Fetal Murine Fancd2-/-Hematopoietic Stem and Progenitor Cells."Published figure using Phospho-p38 MAPK (Thr180, Tyr182) monoclonal antibody (Product # 46-9078-42) in Flow Cytometry"

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