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產(chǎn)品訂購信息：
英文名稱  Anti-phospho-PAK1(Thr212) 

中文名稱   磷酸化p21激活激酶1抗體說明書 

別    名  PAK1(Phospho-Thr212); PAK1(phospho T212); Alpha PAK; Alpha-PAK; MGC130000; MGC130001; p21 activated kinase 1; p21 protein (Cdc42/Rac) activated kinase 1; p21-activated kinase 1; p21/Cdc42/Rac1 activated kinase 1 (yeast Ste20 related); p21/Cdc42/Rac1 activated kinase 1; p21/Cdc42/Rac1-activated kinase 1 (STE20 homolog, yeast); p65 PAK; p65-PAK; PAK alpha; PAK-1; Pak1; PAK1_HUMAN; Paka; PAKalpha; Protein kinase MUK2; Rac/p21-activated kinase; Serine/threonine-protein kinase PAK 1; STE20 homolog (yeast); STE20 homolog yeast.

濃    度   1mg/1ml

規(guī) 格  0.1ml/100μg

抗體來源   Rabbit

克隆類型   polyclonal

交叉反應(yīng)   Human, Mouse, Rat, Dog, Rabbit 

產(chǎn)品類型   一抗 磷酸化抗體  

研究領(lǐng)域     免疫學(xué) 信號轉(zhuǎn)導(dǎo) 轉(zhuǎn)錄調(diào)節(jié)因子 激酶和磷酸酶

蛋白分子量  predicted molecular weight: 60kDa

性    狀   Lyophilized or Liquid

免 疫 原 KLH conjugated Synthesised phosphopeptide derived from human PAK1 around the phosphorylation site of Thr212 

亞    型   IgG

純化方法   affinity purified by Protein A

儲 存 液   0.01M PBS, pH 7.4 with 10 mg/ml BSA and 0.1% Sodium azide

磷酸化p21激活激酶1抗體說明書 產(chǎn)品應(yīng)用    WB=1:100-500 ELISA=1:500-1000 IP=1:20-100 IHC-P=1:100-500 IHC-F=1:100-500 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.  

保存條件  Store at -20 °C for one year. Avoid repeated freeze/thaw cycles. The lyophilized antibody is stable at room temperature for at least one month and for greater than a year when kept at -20°C. When reconstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4 °C. 

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

產(chǎn)品介紹 The p21 activated kinases (PAK) are critical effectors that link Rho GTPases to cytoskeleton reorganization and nuclear signaling. The PAK proteins are a family of serine/threonine kinases that serve as targets for the small GTP binding proteins, CDC42 and RAC1, and have been implicated in a wide range of biological activities. The protein encoded by this gene is activated by proteolytic cleavage during caspase-mediated apoptosis, and may play a role in regulating the apoptotic events in the dying cell.

Function : The activated kinase acts on a variety of targets. Likely to be the GTPase effector that links the Rho-related GTPases to the JNK MAP kinase pathway. Activated by CDC42 and RAC1. Involved in dissolution of stress fibers and reorganization of focal complexes. Involved in regulation of microtubule biogenesis through phosphorylation of TBCB. DVL1 and PAK1 form a ternary complex with MUSK which is important for MUSK-dependent regulation of AChR clustering during the formation of the neuromuscular junction (NMJ). Activity is inhibited in cells undergoing apoptosis, potentially due to binding of CDC2L1 and CDC2L2. Phosphorylates MYL9/MLC2. Phosphorylates RAF1 at 'Ser-338' and 'Ser-339' resulting in: activation of RAF1, stimulation of RAF1 translocation to mitochondria, phosphorylation of BAD by RAF1, and RAF1 binding to BCL2.

Subunit : Homodimer in its autoinhibited state. Active as monomer. Interacts tightly with GTP-bound but not GDP-bound CDC42/P21 and RAC1. Binds to the caspase-cleaved p110 isoform of CDC2L1 and CDC2L2, p110C, but not the full-length proteins. Component of cytoplasmic complexes, which also contain PXN, ARHGEF6 and GIT1. Interacts with ARHGEF7. Also interacts with CRIPAK. Interacts with NISCH. Interacts with DVL1; mediates the formation of a DVL1, MUSK and PAK1 ternary complex involved in AChR clustering. Probably found in a ternary complex composed of DSCAM, PAK1 and RAC1. Interacts with DSCAM (via cytoplasmic domain); the interaction is direct and enhanced in presence of RAC1. Interacts with SCRIB. Interacts with PDPK1. Interacts (via kinase domain) with RAF1. Interaction with NCK1 and NCK2.

Subcellular Location : Cytoplasm. Cell junction, focal adhesion. Note=Recruited to focal adhesions upon activation.

Similarity : Belongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. STE20 subfamily.

Contains 1 CRIB domain.

Contains 1 protein kinase domain.

Database links : UniProtKB/Swiss-Prot: Q13153.2

PAK1-3蛋白具有廣泛的生物學(xué)功能,也是一個保守的絲氨酸/蘇氨酸蛋白激酶,參與許多重要的細胞活動。包括細胞骨架的動力學(xué)調(diào)節(jié),細胞移動,生存和凋亡,細胞周期,基因轉(zhuǎn)錄調(diào)節(jié),細胞生長信號轉(zhuǎn)導(dǎo)和轉(zhuǎn)化等。

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抗體的生物素化標記實驗要點：

1. 磷酸化p21激活激酶1抗體說明書 如在反應(yīng)混合液中有疊氮鈉或游離氨基存在,會抑制標記反應(yīng)。因此,蛋白質(zhì)在反應(yīng)前要對 0.1mol/L碳酸氫鈉緩沖液或0.5mol/L硼酸緩沖液充分透析；

2.所用的NHSB及待生物素化蛋白質(zhì)之間的分子比按蛋白質(zhì)表面的ε-氨基的密度會有所不同,選擇不當(dāng)則影響標記的效率,應(yīng)先用幾個不同的分子比來篩選最適條件；

3.用NHSB量過量也是不利的,抗原的結(jié)合位點可能因此被封閉,導(dǎo)致抗體失活；

4.由于抗體的氨基不易接近可能造成生物素化不足,此時可加入去污劑如 Triton x-100, Tween20等；

5.當(dāng)游離ε-氨基(賴氨酸殘基的氨基)存在于抗體的抗原結(jié)合位點時,或位于酶的催化位點時,生物素化會降低或損傷抗體蛋白的結(jié)合力或活性；

6.生物素還可能與不同的功能基團,如羰基、氨基、巰基、異咪唑基及苯酚基,也可與糖基共價結(jié)合；

7.交聯(lián)反應(yīng)后,應(yīng)充分透析,否則,殘余的生物素會對生物素化抗體與親和素的結(jié)合產(chǎn)生競爭作用；

8.在細胞的熒光標記實驗中,中和親和素的本底低,但由于鏈霉親和素含有少量正電荷,故對某些細胞可導(dǎo)致高本底。

抗體的鑒定：

1）磷酸化p21激活激酶1抗體說明書 抗體的效價鑒定：不管是用于診斷還是用于,制備抗體的目的都是要求較高效價。不同的抗原制備的抗體,要求的效價不一。鑒定效價的方法很多,包括有試管凝集反應(yīng),瓊脂擴散試驗,酶聯(lián)免疫吸附試驗等。常用的抗原所制備的抗體一般都有約成的鑒定效價的方法,以資比較。如制備抗抗體的效價,一般就采用瓊脂擴散試驗來鑒定。

2）抗體的特異性鑒定：抗體的特異性是指與相應(yīng)抗原或近似抗原物質(zhì)的識別能力。抗體的特異性高,它的識別能力就強。衡量特異性通常以交叉反應(yīng)率來表示。交叉反應(yīng)率可用競爭抑制試驗測定。以不同濃度抗原和近似抗原分別做競爭抑制曲線,計算各自的結(jié)合率,求出各自在IC50時的濃度,并按公式計算交叉反應(yīng)率。 

如果所用抗原濃度IC50濃度為pg/管,而一些近似抗原物質(zhì)的IC50濃度幾乎是無窮大時,表示這一抗血清與其他抗原物質(zhì)的交叉反應(yīng)率近似為0,即該血清的特異性較好。

3）抗體親和力：是指抗體和抗原結(jié)合的牢固程度。親和力的高低是由抗原分子的大小,抗體分子的結(jié)合位點與抗原決定簇之間立體構(gòu)型的合適度決定的。有助于維持抗原抗體復(fù)合物穩(wěn)定的分子間力有氫鍵,疏水鍵,側(cè)鏈相反電荷基因的庫侖力,范德華力和空間斥力。親和力常以親和常數(shù)K表示,K的單位是L/mol。抗體親和力的測定對抗體的篩選,確定抗體的用途,驗證抗體的均一性等均有重要意義。