| 產品編號 | bs-3162R |
| 英文名稱 | Rabbit Anti-Phospho-FAK (Tyr576 + Tyr577) antibody |
| 中文名稱 | 磷酸化粘著斑激酶抗體 |
| 別 名 | FAK (phospho Y576 + Y577); p-FAK (phospho Y576 + Y577); FAK (phospho-Tyr576/Tyr577); FADK 1; FADK; FAK 1; FAK related non kinase polypeptide; FAK1; Focal adhesion kinase 1; FRNK; pp125FAK; Protein tyrosine kinase 2; Protein Tyrosine Kinase Cytoplasmic; PTK 2; FAK1_HUMAN; Focal adhesion kinase-related nonkinase; Protein phosphatase 1 regulatory subunit 71; PPP1R71; Protein-tyrosine kinase 2; p125FAK. |
| 產品類型 | 磷酸化抗體 |
| 研究領域 | 激酶和磷酸酶 |
| 抗體來源 | Rabbit |
| 克隆類型 | Polyclonal |
| 交叉反應 | Human,Mouse,Rat (predicted: Rabbit,Cow,Chicken,Dog,Horse) |
| 產品應用 | WB=1:500-2000,IHC-P=1:100-500,IHC-F=1:100-500,Flow-Cyt=1ug/Test,IF=1:100-500,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 | 116kDa |
| 細胞定位 | 細胞核 細胞漿 細胞膜 |
| 性 狀 | Liquid |
| 濃 度 | 1mg/ml |
| 免 疫 原 | KLH conjugated Synthesised phosphopeptide derived from human FAK around the phosphorylation site of Tyr576/577: ST(p-Y)(p-Y)KA |
| 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 |
Non-receptor protein-tyrosine kinase implicated in signaling pathways involved in cell motility, proliferation and apoptosis. Activated by tyrosine-phosphorylation in response to either integrin clustering induced by cell adhesion or antibody cross-linking, or via G-protein coupled receptor (GPCR) occupancy by ligands such as bombesin or lysophosphatidic acid, or via LDL receptor occupancy. Plays a potential role in oncogenic transformations resulting in increased kinase activity. Function: Non-receptor protein-tyrosine kinase implicated in signaling pathways involved in cell motility, proliferation and apoptosis. Activated by tyrosine-phosphorylation in response to either integrin clustering induced by cell adhesion or antibody cross-linking, or via G-protein coupled receptor (GPCR) occupancy by ligands such as bombesin or lysophosphatidic acid, or via LDL receptor occupancy. Microtubule-induced dephosphorylation at Tyr-397 is crucial for the induction of focal adhesion disassembly. Plays a potential role in oncogenic transformations resulting in increased kinase activity. Subunit: Interacts (via first Pro-rich region) with CAS family members (via SH3 domain), including BCAR1, BCAR3, CASS4 and NEDD9. Interacts with GIT1. Interacts with SORBS1. Interacts with RGNEF. Interacts with SHB. Interacts with PXN and TLN1. Interacts with STAT1. Interacts with DCC. Interacts with WASL. Interacts with ARHGEF7. Interacts with GRB2 and GRB7 (By similarity). Component of a complex that contains at least FER, CTTN and PTK2/FAK1. Interacts with BMX. Interacts with TGFB1I1. Interacts with STEAP4. Interacts with ZFYVE21. Interacts with ESR1. Interacts with PIK3R1 or PIK3R2. Interacts with SRC, FGR, FLT4 and RET. Interacts with EPHA2 in resting cells; activation of EPHA2 recruits PTPN11, leading to dephosphorylation of PTK2/FAK1 and dissociation of the complex. Interacts with EPHA1 (kinase activity-dependent). Interacts with CD4; this interaction requires the presence of HIV-1 gp120. Interacts with PIAS1. Interacts with ARHGAP26 and SHC1. Interacts with RB1CC1; this inhibits PTK2/FAK1 activity and activation of downstream signaling pathways. Interacts with P53/TP53 and MDM2. Interacts with LPXN (via LD motif 3). Subcellular Location: Cell junction, focal adhesion. Cell membrane; Peripheral membrane protein; Cytoplasmic side. Cytoplasm, cell cortex. Cytoplasm, cytoskeleton. Cytoplasm, cytoskeleton, centrosome. Nucleus. Note=Constituent of focal adhesions. Detected at microtubules. Tissue Specificity: Detected in B and T-lymphocytes. Isoform 1 and isoform 6 are detected in lung fibroblasts (at protein level). Ubiquitous. Post-translational modifications: Phosphorylated on tyrosine residues upon activation, e.g. upon integrin signaling. Tyr-397 is the major autophosphorylation site, but other kinases can also phosphorylate this residue. Phosphorylation at Tyr-397 promotes interaction with SRC and SRC family members, leading to phosphorylation at Tyr-576, Tyr-577 and at additional tyrosine residues. FGR promotes phosphorylation at Tyr-397 and Tyr-576. FER promotes phosphorylation at Tyr-577, Tyr-861 and Tyr-925, even when cells are not adherent. Tyr-397, Tyr-576 and Ser-722 are phosphorylated only when cells are adherent. Phosphorylation at Tyr-397 is important for interaction with BMX, PIK3R1 and SHC1. Phosphorylation at Tyr-925 is important for interaction with GRB2. Dephosphorylated by PTPN11; PTPN11 is recruited to PTK2 via EPHA2 (tyrosine phosphorylated). Microtubule-induced dephosphorylation at Tyr-397 is crucial for the induction of focal adhesion disassembly; this dephosphorylation could be catalyzed by PTPN11 and regulated by ZFYVE21. Sumoylated; this enhances autophosphorylation. DISEASE: Note=Aberrant PTK2/FAK1 expression may play a role in cancer cell proliferation, migration and invasion, in tumor formation and metastasis. PTK2/FAK1 overexpression is seen in many types of cancer. Similarity: Belongs to the protein kinase superfamily. Tyr protein kinase family. FAK subfamily. Contains 1 FERM domain. SWISS: Q05397 Gene ID: 5747 Database links: Entrez Gene: 5747 Human Entrez Gene: 14083 Mouse Omim: 600758 Human SwissProt: Q05397 Human SwissProt: P34152 Mouse Unigene: 395482 Human Unigene: 254494 Mouse Unigene: 2809 Rat FAK是整合蛋白介導的信號轉導中的重要成員,有酪氨酸蛋白激酶活性,并可自身磷酸化,FAK本身是胱冬肽酶(caspase)的底物。作為信號分子的FAK參與抑制細胞凋亡并直接參與細胞多種功能的調節。 1.FAK 局部粘著斑激酶，是一種酪氨酸激酶;腫瘤細胞的侵襲性生長是一個多步驟的復雜過程，有多種生物化學因子參與其中,局部粘著斑激酶（focal adhesion kinase, FAK）介導的信號轉導系統就是其中最為重要的細胞信號轉導途徑之一。腫瘤細胞必須黏附于細胞外基質，通過促進依賴于PTK激酶活性的細胞外基質信號轉導，進而影響細胞的黏附、運動與遷移。 2.粘著斑激酶(focal adhesion kinase,FAK)是整合蛋白介導的信號轉導中的重要成員,有酪氨酸蛋白激酶活性,并可自身磷酸化；為信號分子的FAK,還與細胞內其他信號轉導通路存在串話(crosstalk),直接參與了細胞多種功能的調節。 3.盡管FAK的確切功能尚不清楚,但若干實驗均提示FAK可能有兩個作用,一是在細胞鋪展和移動時,FAK參與粘著斑形成和調節；二是FAK參與信號轉導過程,以告知細胞核其細胞已錨定了。近年有關FAK在細胞凋亡中的作用也業已肯定。 |
| 產品圖片 |
Sample:
B16(Mouse) Cell Lysate at 40 ug
Primary: Anti-Phospho-FAK(Tyr576 + Tyr577) (bs-3162R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 116 kD
Observed band size: 116 kD
Sample:
HT1080(Human) Cell Lysate at 30 ug
Hela(Human) Cell Lysate at 30 ug
HepG2(Human) Cell Lysate at 30 ug
Primary: Anti-Phospho-FAK(Tyr576 + Tyr577) (bs-3162R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 116 kD
Observed band size: 116 kD
Paraformaldehyde-fixed, paraffin embedded (rat testis); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Phospho-FAK (Tyr576 + Tyr577)) Polyclonal Antibody, Unconjugated (bs-3162R ) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Phospho-FAK (Tyr576 + Tyr577)) Polyclonal Antibody, Unconjugated (bs-3162R ) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Phospho-FAK (Tyr576 + Tyr577)) Polyclonal Antibody, Unconjugated (bs-3162R ) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Phospho-FAK (Tyr576 + Tyr577)) Polyclonal Antibody, Unconjugated (bs-3162R ) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Tissue/cell: human pneumonia 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-FAK(Tyr576/577) Polyclonal Antibody, Unconjugated(bs-3162R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
Blank control:A431.
Primary Antibody (green line): Rabbit Anti-Phospho-FAK (Tyr576 + Tyr577) antibody (bs-3162R)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-AF647
Dilution: 1μg /test.
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.
Blank control: A431.
Primary Antibody (green line): Rabbit Anti-Phospho-FAK (Tyr576 + Tyr577) antibody (bs-3162R)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-AF647
Dilution: 1μg /test.
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.
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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 | |
