Invitrogen

Phospho-p38 MAPK (Thr180, Tyr182) Monoclonal Antibody (4NIT4KK), PE-Cyanine7, eBioscience™

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说明书
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说明书
实验方案
问题与答案
技术支持

产品信息

25-9078-42

应用
建议稀释比
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流式细胞分析 (Flow)

5 µL (0.25 µg)/test
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产品规格

种属反应

Human, Mouse

宿主/亚型

Mouse / IgG2b, kappa

推荐的同型对照

Mouse IgG2b kappa Isotype Control (eBMG2b), PE-Cyanine7, eBioscience™

分类

Monoclonal

类型

Antibody

克隆号

4NIT4KK

偶联物

PE-Cyanine7 PE-Cyanine7 PE-Cyanine7

激发/发射光谱

569/780 nm 查看光谱 spectra

形式

Liquid

浓度

5 µL/Test

纯化类型

Affinity chromatography

保存液

PBS, pH 7.2, with BSA

内含物

0.09% sodium azide

保存条件

4°C, store in dark, DO NOT FREEZE!

运输条件

Ambient (domestic); Wet ice (international)

RRID

AB_2802267

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-diluted and tested by intracellular staining followed by flow cytometric analysis of stimulated normal human peripheral blood cells. This may be used at 5 µL (0.25 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 BestProtocols 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 BestProtocols® 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 (Product # 00-8222-49) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-57) 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.

靶标信息

The p38 MAPK cascade regulates a variety of cellular responses to stress, inflammation, and other signals. p38 MAPK is relatively inactive in the nonphosphorylated form and becomes rapidly activated by dual phosphorylation of a Thr-Gly-Tyr motifs. There are four isoforms of p38 MAPK, which differ in their tissue expression and affinity for upstream activators and downstream effectors. When cells are exposed to tumor necrosis factor, interleukin-1, heat shock, or other activating stimuli, activation of MAPK kinase-3 occurs by phosphorylation. Activated MAPK kinase-3/6 phosphorylate each residue of Thr180 and Tyr182 in p38 MAPK. Phospho-p38 MAPK activates ATF-2, CHOP-1, MEF-2 and other transcription factors through phosphorylation.

仅用于科研。不用于诊断过程。未经明确授权不得转售。

How to use the Panel Builder

Watch the video to learn how to use the Invitrogen Flow Cytometry Panel Builder to build your next flow cytometry panel in 5 easy steps.

生物信息学

蛋白别名: CRK1; CSAID-binding protein; Csaids binding protein; cytokine suppressive anti-inflammatory drug binding protein; cytokine suppressive anti-inflammatory drug binding protein 1; Cytokine suppressive anti-inflammatory drug-binding protein; MAP kinase 14; MAP kinase MXI2; MAP kinase p38 alpha; MAPK 14; MAX-interacting protein 2; mitogen activated protein kinase 14; Mitogen-activated protein kinase 14; Mitogen-activated protein kinase p38 alpha; p38 alpha; p38 MAP kinase; p38 MAP kinase alpha; p38 MAPK; p38 mitogen activated protein kinase; p38alpha Exip; SAPK2a; Stress-activated protein kinase 2a; tRNA synthetase cofactor p38

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基因别名: Crk1; CSBP; CSBP1; CSBP2; CSPB1; EXIP; MAPK14; MXI2; p38; p38-alpha; p38a; p38ALPHA; p38MAPK; PRKM14; PRKM15; RK; SAPK2A

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UniProt ID: (Human) Q16539, (Mouse) P47811

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Entrez Gene ID: (Human) 1432, (Mouse) 26416

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功能
protein serine/threonine kinase activity MAP kinase activity MAP kinase kinase activity protein binding ATP binding protein C-terminus binding enzyme binding protein phosphatase binding NFAT protein binding nucleotide binding protein kinase activity kinase activity transferase activity transferase activity, transferring phosphorus-containing groups non-receptor serine/threonine protein kinase
参与通路
DNA damage checkpoint activation of MAPK activity cell morphogenesis cartilage condensation angiogenesis placenta development chondrocyte differentiation glucose metabolic process transcription, DNA-templated regulation of transcription from RNA polymerase II promoter apoptotic process movement of cell or subcellular component chemotaxis signal transduction cell surface receptor signaling pathway transmembrane receptor protein serine/threonine kinase signaling pathway Ras protein signal transduction skeletal muscle tissue development response to glucose positive regulation of gene expression positive regulation of myotube differentiation myoblast differentiation involved in skeletal muscle regeneration peptidyl-serine phosphorylation fatty acid oxidation osteoclast differentiation positive regulation of cyclase activity lipopolysaccharide-mediated signaling pathway response to muramyl dipeptide intracellular signal transduction cellular response to vascular endothelial growth factor stimulus response to muscle stretch p38MAPK cascade positive regulation of protein import into nucleus signal transduction in response to DNA damage positive regulation of blood vessel endothelial cell migration positive regulation of erythrocyte differentiation positive regulation of myoblast differentiation positive regulation of transcription from RNA polymerase II promoter positive regulation of glucose import protein autophosphorylation vascular endothelial growth factor receptor signaling pathway regulation of sequence-specific DNA binding transcription factor activity striated muscle cell differentiation positive regulation of muscle cell differentiation positive regulation of cardiac muscle cell proliferation 3'-UTR-mediated mRNA stabilization cellular response to lipopolysaccharide cellular response to ionizing radiation negative regulation of canonical Wnt signaling pathway positive regulation of brown fat cell differentiation stress-induced premature senescence cellular response to virus regulation of cytokine production involved in inflammatory response positive regulation of myoblast fusion regulation of signal transduction by p53 class mediator positive regulation of reactive oxygen species metabolic process positive regulation of interleukin-12 secretion regulation of transcription, DNA-templated protein phosphorylation cellular response to DNA damage stimulus phosphorylation response to lipopolysaccharide stress-activated MAPK cascade
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