?5300?????? PathScan? Bcr/Abl Activity Assay: Phospho-c-Abl, Phospho-Stat5 and Phospho-CrkL Multiplex Western Detection Cocktail

5301?????? PathScan? Multiplex Western Cocktail I: Phospho-p90RSK, Phospho-Akt, Phospho-p44/42 MAPK (Erk1/2) and Phospho-S6 Ribosomal Protein Detection Cocktail I

5302?????? PathScan? Multiplex Western Cocktail II: Phospho-p90RSK, Phospho-p53, Phospho-p38 MAPK and Phospho-S6 Ribosomal Protein Detection Cocktail II

5303?????? PathScan? Multiplex Western Cocktail III: Phospho-Stat1, Phospho-SAPK/JNK, Phospho-S6 Ribosomal Protein and Phospho-HSP27 Detection Cocktail III

5304?????? PathScan? PDGFR Activity Assay: Phospho-PDGFR, Phospho-SHP2, Phospho-Akt, and Phospho-p44/42 MAPK (Erk1/2) Multiplex Western Detection Cocktail II

該cocktail中的每個磷酸化抗體都能識別內源性特異靶點磷酸化的蛋白水平。eIF4E Antibody都能檢測它的總靶蛋白水平，并且不依賴于磷酸化，同時該抗體還能用于蛋白負載對照。

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Source / Purification

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Polyclonal antibodies are produced by immunizing animals with synthetic peptides. Antibodies are purified by protein A and peptide affinity chromatography.

該多克隆抗體通過使用合成肽段免疫動物而獲得。該抗體經蛋白A和肽親和層析純化。

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Description

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The PathScan? Multiplex Western Cocktail II offers a unique method to assay the activation of multiple pathways on one membrane without stripping and reprobing. This method saves the user valuable time, while increasing accuracy and minimizing reagent waste. The system allows the user to simultaneously detect levels of phospho-p90RSK, phospho-p53, phospho-p38 MAPK and phospho-S6 ribosomal protein. The kit also includes elF4E antibody to control protein loading.

PathScan? Multiplex Western Cocktail IIPathscan? Multiplex Western Cocktail II為分析一張膜上各種蛋白的活性狀態提供了獨特的方法，不用去除免疫印跡膜上的封閉液和抗體，然后再次重新標記。該方法為使用者節約了時間，且調高了準確度，減少了試劑的浪費。該系統讓使用者能即時檢測phospho-p90RSK, phospho-p53, phospho-p38 MAPK和phospho-S6核糖體蛋白。該試劑盒也包括用于控制蛋白負載的elF4E Antibody。

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Western Blotting

Western blot analysis of extracts from Mv1Lu mink lung epithelial cells untreated or treated with UV and TPA, using PathScan Multiplex Western cocktail II to detect phosphorylation of p90RSK, p53, p38 MAPK and S6 ribosomal protein.Western blot方法檢測細胞提取物：未經處理和UV、TPA處理的Mv1Lu貂肺上皮細胞。使用的抗體是PathScan Multiplex Western cocktail II，以檢測p90RSK, p53, p38 MAPK、S6核糖體蛋白的磷酸化。

Western Blotting

Western blot analysis of extracts from Mv1Lu mink lung epithelial cells untreated or treated with UV and TPA, using PathScan Multiplex Western cocktail II to detect phosphorylation of p90RSK, p53, p38 MAPK and S6 ribosomal protein.

Background

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The 90 kDa ribosomal S6 kinases (RSK1-3) are a family of serine/threonine kinases broadly expressed in response to many growth factors, polypeptide hormones and neurotransmitters (1). p90RSK is activated by Erk1 and Erk2 in vitro and in vivo via phosphorylation (2). Several sites, such as Ser380, Thr359 and Ser363, are important for its activation (3). ? The p53 tumor suppressor protein plays a major role in cellular response to DNA damage and other genomic aberrations. Activation of p53 can lead to either cell cycle arrest and DNA repair or apoptosis (4). p53 is phosphorylated at multiple sites in vivo and by several different protein kinases in vitro (5,6). DNA damage induces phosphorylation of p53 at Ser15 and Ser20 and leads to reduced interaction of p53 with its negative regulator, oncoprotein MDM2 (7). ? p38 MAP kinase controls cellular responses to cytokines and stress (8-11). Like the SAPK/JNK pathway, p38 MAP kinase is activated by a variety of cellular stresses including osmotic shock, inflammatory cytokines, lipopolysaccharides (LPS), UV light and growth factors (8-12). MKK3, MKK6 and SEK activate p38 MAP kinase by phosphorylation at Thr180 and Tyr182. ? Growth factors and mitogens induce the activation of p70 S6 kinase, which in turn phosphorylates the S6 ribosomal protein. Phosphorylation of S6 correlates with an increase in translation, particularly of mRNAs with an oligopyrimidine tract in their 5' untranslated regions (13). This group of mRNAs (5'TOP) encodes proteins involved in cell cycle progression and proteins that are part of the translational machinery, such as ribosomal proteins and elongation factors (13,14).

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90 kDa核糖體S6激酶(RSK1-3)是一類絲氨酸/蘇氨酸激酶家族，廣泛表達于許多生長因子、多肽激素和神經傳導物質的應答反應之后(1)。p90RSK在體外能被Erk1、Erk2能激活，在體內能通過磷酸化而激活(2)。一些位點如絲氨酸（380、363位點）、蘇氨酸（359位點），它的活化十分重要(3)。p53腫瘤抑制子蛋白在細胞應答DNA損傷和其他基因組紊亂中起著主要作用。p53的活化導致細胞周期捕獲和DNA修復或凋亡(4)。p53在體內許多位點被磷酸化，在體外被許多不同的蛋白激酶磷酸化(5,6)。DNA損傷誘導p53絲氨酸（15、20位點）的磷酸化，從而導致p53與它的負調節子——腫瘤蛋白MDM2的相互作用減少(7)。p38 MAP kinase控制細胞對細胞因子和應激的應答反應(8-11)。在SAPK/JNK通路中，p38 MAP kinase被許多細胞應激激活，包括滲透性休克、炎性細胞因子、脂多糖（LPS）、UV照射和生長因子(8-12)。MKK3, MKK6，SEK通過磷酸化p38 MAP kinase蘇氨酸（180位點）和酪氨酸（182位點）而使其活化。生長因子和有絲分裂原誘導 p70 S6 kinase的活化，該激酶又磷酸化S6核糖體蛋白。S6的磷酸化與轉位的提高有關，特別是5' 非轉位區域內帶帽序列的mRNAs(13)。mRNAs (5'TOP)編碼的蛋白涉及細胞周期進程和轉位機制部分的蛋白，如核糖體蛋白和延長因子(13,14)。

11989 Phospho-p90RSK (Ser380) (D3H11) Rabbit mAb

12630 SignalFire? Plus ECL Reagent

12757 SignalFire? Elite ECL Reagent

4511 Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP? Rabbit mAb

4858 Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP? Rabbit mAb

6883 SignalFire? ECL Reagent

7074 Anti-rabbit IgG, HRP-linked Antibody

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7727 Biotinylated Protein Ladder Detection Pack