| 產品編號 | bs-2270R |
| 英文名稱 | Phospho-PAK4 (Ser99) Rabbit pAb |
| 中文名稱 | 磷酸化p21激活激酶4抗體 |
| 別 名 | p21 activated kinase 4; p21(CDKN1A) activated kinase 4; PAK 4; Protein kinase related to S.cerevisiae STE20 effector for Cdc42Hs; Serine threonine kinase PAK 4; Serine/threonine protein kinase PAK 4; Serine/threonine protein kinase PAK4; KIAA1142. |
| 產品類型 | 磷酸化抗體 |
| 研究領域 | 腫瘤 細胞生物 免疫學 信號轉導 細胞凋亡 轉錄調節因子 激酶和磷酸酶 |
| 抗體來源 | Rabbit |
| 克隆類型 | Polyclonal |
| 交叉反應 | Human,Mouse,Rabbit (predicted: Rat,Cow,Chicken,Dog,Horse) |
| 產品應用 | WB=1:500-2000,IHC-P=1:100-500,IHC-F=1:100-500,IF=1:100-500,Flow-Cyt=1ug/Test,ELISA=1:5000-10000
not yet tested in other applications. optimal dilutions/concentrations should be determined by the end user. |
| 理論分子量 | 64 kDa |
| 檢測分子量 | |
| 細胞定位 | 細胞漿 |
| 性 狀 | Liquid |
| 濃 度 | 1mg/ml |
| 免 疫 原 | KLH conjugated Synthesised phosphopeptide derived from human PAK4 around the phosphorylation site of Ser99: SN(p-S)LR |
| 亞 型 | IgG |
| 純化方法 | affinity purified by Protein A |
| 緩 沖 液 | 0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol. |
| 保存條件 | Shipped at 4℃. Store at -20℃ for one year. Avoid repeated freeze/thaw cycles. |
| 注意事項 | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications. |
| PubMed | PubMed |
| 產品介紹 |
p21-activated kinases (PAKs) belong to the family of serine/threonine kinases involved in the control of various cellular processes, including the cell cycle, dynamics of the cytoskeleton, apoptosis, oncogenic transformation, and transcription. All PAK family members are characterized by the presence of p21-binding domain. p21-activated kinases are regulated by the small GTP-binding proteins Rac and Cdc42, and lipids, which stimulate autophosphorylation and phosphorylation of exogenous substrates. Serine (Ser-474) is the likely autophosphorylation site in the kinase domain of PAK4 in vivo. Phosphospecific antibodies directed against serine 474 detect activated PAK4 on the Golgi membrane when PAK4 is co-expressed with activated Cdc42. Current data strongly implicates PAK-4 in oncogenesis. PAK4 is frequently overexpressed in human tumor cell lines of various tissue origins. Function: Serine/threonine protein kinase that plays a role in a variety of different signaling pathways including cytoskeleton regulation, cell migration, growth, proliferation or cell survival. Activation by various effectors including growth factor receptors or active CDC42 and RAC1 results in a conformational change and a subsequent autophosphorylation on several serine and/or threonine residues. Phosphorylates and inactivates the protein phosphatase SSH1, leading to increased inhibitory phosphorylation of the actin binding/depolymerizing factor cofilin. Decreased cofilin activity may lead to stabilization of actin filaments. Phosphorylates LIMK1, a kinase that also inhibits the activity of cofilin. Phosphorylates integrin beta5/ITGB5 and thus regulates cell motility. Phosphorylates ARHGEF2 and activates the downstream target RHOA that plays a role in the regulation of assembly of focal adhesions and actin stress fibers. Stimulates cell survival by phosphorylating the BCL2 antagonist of cell death BAD. Alternatively, inhibits apoptosis by preventing caspase-8 binding to death domain receptors in a kinase independent manner. Plays a role in cell-cycle progression by controlling levels of the cell-cycle regulatory protein CDKN1A and by phosphorylating RAN. Subunit: Interacts with FGFR2 and GRB2. Interacts tightly with GTP-bound but not GDP-bound CDC42/p21 and weakly with RAC1. Subcellular Location: Cytoplasm. Note=Seems to shuttle between cytoplasmic compartments depending on the activating effector. For example, can be found on the cell periphery after activation of growth-factor or integrin-mediated signaling pathways. Tissue Specificity: Highest expression in prostate, testis and colon. Post-translational modifications: Autophosphorylated on serine residues when activated by CDC42/p21. Phosphorylated on tyrosine residues upon stimulation of FGFR2. Similarity: Belongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. STE20 subfamily. Contains 1 CRIB domain. Contains 1 protein kinase domain. SWISS: O96013 Gene ID: 10298 Database links: Entrez Gene: 10298 Human Entrez Gene: 70584 Mouse Omim: 605451 Human SwissProt: O96013 Human SwissProt: Q8BTW9 Mouse Unigene: 20447 Human Unigene: 21876 Mouse Unigene: 18543 Rat 絲氨酸/蘇氨酸蛋白激酶4 (PAK4)是新發現的一類蛋白因子，在細胞信號轉導過程中起很重要的作用。Pak4常在人體的多種組織器官的腫瘤細胞中呈現過表達，他的絲氨酸位點很可能是Pak4酶活性區域中的自身磷酸化位點，絲氨酸到谷氨酸位點的突變體促使其獲得了酶活性。在腫瘤遺傳學、細胞癌變、細胞凋亡等一些重要的細胞生物學研究中，具有重要的意義。 |
| 產品圖片 |
Sample:
Hela(Human) Cell Lysate at 30 ug
Primary: Anti-Phospho-PAK4(Ser99) (bs-2270R) at 1/500 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 64 kD
Observed band size: 64 kD
Sample:
293T(Human) Cell Lysate at 30 ug
Primary: Anti-Phospho-PAK4(Ser99) (bs-2270R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 64 kD
Observed band size: 64 kD
Tissue/cell: human prostate tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-Phospho-PAK4 (Ser99) Polyclonal Antibody, Unconjugated(bs-2270R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
Blank control（black line）:NIH/3T3.
Primary Antibody (green line): Rabbit Anti-Phospho-PAK4 (Ser99) antibody (bs-2270R)
Dilution:1ug/Test;
Secondary Antibody（white blue line）: Goat anti-rabbit IgG-AF488
Dilution: 0.5ug/Test.
Isotype control（orange line）: Normal Rabbit IgG
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at -20℃, The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
phosphopeptide
non phosphopeptide
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| 1、抗體溶解方法 | |
| 2、抗體修復方式 | |
| 3、常用試劑的配制 | |
| 4、免疫組化操作步驟 | |
| 5、免疫組化問題解答 | |
| 6、Western Blotting 操作步驟 | |
| 7、Western Blotting 問題解答 | |
| 8、關于肽鏈的設計 | |
| 9、多肽的溶解與保存 | |
| 10、酶標抗體效價測定程序 | |
