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粘著斑激酶抗體

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中文名稱 粘著斑激酶抗體
別    名 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.

 

研究領(lǐng)域 腫瘤  細(xì)胞生物  免疫學(xué)  激酶和磷酸酶  
抗體來源 Rabbit
克隆類型 Polyclonal
交叉反應(yīng) Human, Mouse, Rat, 
產(chǎn)品應(yīng)用 WB=1:500-2000 ELISA=1:500-1000 IHC-P=1:100-500 IHC-F=1:100-500 Flow-Cyt=1μg/Test IF=1:100-500 (石蠟切片需做抗原修復(fù))
not yet tested in other applications.
optimal dilutions/concentrations should be determined by the end user.
分 子 量 116kDa
細(xì)胞定位 細(xì)胞核 細(xì)胞漿 細(xì)胞膜 
性    狀 Liquid
濃    度 1mg/ml
免 疫 原 KLH conjugated synthetic peptide derived from human FAK:901-1052/1052 
亞    型 IgG
純化方法 affinity purified by Protein A
儲 存 液 0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol.
保存條件 Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.
PubMed PubMed
產(chǎn)品介紹 This gene encodes a cytoplasmic protein tyrosine kinase which is found concentrated in the focal adhesions that form between cells growing in the presence of extracellular matrix constituents. The encoded protein is a member of the FAK subfamily of protein tyrosine kinases but lacks significant sequence similarity to kinases from other subfamilies. Activation of this gene may be an important early step in cell growth and intracellular signal transduction pathways triggered in response to certain neural peptides or to cell interactions with the extracellular matrix. Several transcript variants encoding different isoforms have been found for this gene, but the full-length natures of only three of them have been determined. [provided by RefSeq, Dec 2010]

Function:
Non-receptor protein-tyrosine kinase that plays an essential role in regulating cell migration, adhesion, spreading, reorganization of the actin cytoskeleton, formation and disassembly of focal adhesions and cell protrusions, cell cycle progression, cell proliferation and apoptosis. Required for early embryonic development and placenta development. Required for embryonic angiogenesis, normal cardiomyocyte migration and proliferation, and normal heart development. Regulates axon growth and neuronal cell migration, axon branching and synapse formation; required for normal development of the nervous system. Plays a role in osteogenesis and differentiation of osteoblasts. Functions in integrin signal transduction, but also in signaling downstream of numerous growth factor receptors, G-protein coupled receptors (GPCR), EPHA2, netrin receptors and LDL receptors. Forms multisubunit signaling complexes with SRC and SRC family members upon activation; this leads to the phosphorylation of additional tyrosine residues, creating binding sites for scaffold proteins, effectors and substrates. Regulates numerous signaling pathways. Promotes activation of phosphatidylinositol 3-kinase and the AKT1 signaling cascade. Promotes activation of MAPK1/ERK2, MAPK3/ERK1 and the MAP kinase signaling cascade. Promotes localized and transient activation of guanine nucleotide exchange factors (GEFs) and GTPase-activating proteins (GAPs), and thereby modulates the activity of Rho family GTPases. Signaling via CAS family members mediates activation of RAC1. Recruits the ubiquitin ligase MDM2 to P53/TP53 in the nucleus, and thereby regulates P53/TP53 activity, P53/TP53 ubiquitination and proteasomal degradation. Phosphorylates SRC; this increases SRC kinase activity. Phosphorylates ACTN1, ARHGEF7, GRB7, RET and WASL. Promotes phosphorylation of PXN and STAT1; most likely PXN and STAT1 are phosphorylated by a SRC family kinase that is recruited to autophosphorylated PTK2/FAK1, rather than by PTK2/FAK1 itself. Promotes phosphorylation of BCAR1; GIT2 and SHC1; this requires both SRC and PTK2/FAK1. Promotes phosphorylation of BMX and PIK3R1. Isoform 6 (FRNK) does not contain a kinase domain and inhibits PTK2/FAK1 phosphorylation and signaling. Its enhanced expression can attenuate the nuclear accumulation of LPXN and limit its ability to enhance serum response factor (SRF)-dependent gene transcription.

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. 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

Entrez Gene: 25614 Rat

Omim: 600758 Human

SwissProt: Q05397 Human

SwissProt: P34152 Mouse

SwissProt: O35346 Rat

Unigene: 395482 Human

Unigene: 254494 Mouse

Unigene: 2809 Rat



Important Note:
This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.

    FAK是整合蛋白介導(dǎo)的信號轉(zhuǎn)導(dǎo)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(xì)胞凋亡并直接參與細(xì)胞多種功能的調(diào)節(jié)。
    1.FAK 局部粘著斑激酶,是一種酪氨酸激酶;腫瘤細(xì)胞的侵襲性生長是一個(gè)多步驟的復(fù)雜過程,有多種生物化學(xué)因子參與其中,局部粘著斑激酶(focal adhesion kinase, FAK)介導(dǎo)的信號轉(zhuǎn)導(dǎo)系統(tǒng)就是其中最為重要的細(xì)胞信號轉(zhuǎn)導(dǎo)途徑之一。腫瘤細(xì)胞必須黏附于細(xì)胞外基質(zhì),通過促進(jìn)依賴于PTK激酶活性的細(xì)胞外基質(zhì)信號轉(zhuǎn)導(dǎo),進(jìn)而影響細(xì)胞的黏附、運(yùn)動(dòng)與遷移。
    2.粘著斑激酶(focal adhesion kinase,FAK)是整合蛋白介導(dǎo)的信號轉(zhuǎn)導(dǎo)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(xì)胞內(nèi)其他信號轉(zhuǎn)導(dǎo)通路存在串話(crosstalk),直接參與了細(xì)胞多種功能的調(diào)節(jié)。
    3.盡管FAK的確切功能尚不清楚,但若干實(shí)驗(yàn)均提示FAK可能有兩個(gè)作用,一是在細(xì)胞鋪展和移動(dòng)時(shí),FAK參與粘著斑形成和調(diào)節(jié);二是FAK參與信號轉(zhuǎn)導(dǎo)過程,以告知細(xì)胞核其細(xì)胞已錨定了。近年有關(guān)FAK在細(xì)胞凋亡中的作用也業(yè)已肯定。
產(chǎn)品圖片 Sample:
MCF-7(Human) Cell Lysate at 30 ug
Primary: Anti-FAK (bs-1340R) 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:
Lane 1: Cerebrum (Mouse) Lysate at 40 ug
Lane 2: NIH/3T3 (Mouse) Cell Lysate at 30 ug
Lane 3: Cerebrum (Rat) Lysate at 40 ug
Lane 4: 293T (Human) Cell Lysate at 30 ug
Lane 5: A431 (Human) Cell Lysate at 30 ug
Lane 6: Huvec (Human) Cell Lysate at 30 ug
Lane 7: A549 (Human) Cell Lysate at 30 ug
Primary: Anti-FAK (bs-1340R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 125 kD
Observed band size: 120 kD
Sample:Thymus (Mouse) Lysate at 40 ug
Primary: Anti-FAK(bs-1340R)at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 116kD
Observed band size: 116kD
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 (FAK) Polyclonal Antibody, Unconjugated (bs-1340R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.Tissue/cell: mouse lung tissue;4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-FAK/PTK2 Polyclonal Antibody, Unconjugated(bs-1340R) 1:200, overnight at 4°C; The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated(bs-0295G-Cy3)used at 1:200 dilution for 40 minutes at 37°C. DAPI(5ug/ml,blue,C-0033) was used to stain the cell nuclei
Blank control (blue line): Hep G2 (blue).
Primary Antibody (green line): Rabbit Anti-FAK antibody (bs-1340R)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE
Dilution: 1μg /test.
Protocol
The cells were fixed with 70% ethanol (Overmight at 4℃) and then permeabilized with 90% ice-cold methanol for 30 min at -20℃.Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2%BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 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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