產(chǎn)品訂購信息：
英文名稱  Anti-Phospho-PAK1(Thr423)/PAK2 (Thr402) 

中文名稱   磷酸化p21激活激酶1/2抗體費用 

別    名  Pak2(phospho T402); Pak1(phospho T423); PAK1(phospho T402); PAK2(phospho T423); PAKalpha; serine/threonine-protein kinase PAK 1; p21 activated kinase 1; PAK alpha; Paka; PAKalpha; Protein kinase MUK2; STE20 homolog (yeast); Alpha PAK; p21/Cdc42/Rac1 activated kinase 1; CB422; EC 2.7.11.1; Gamma PAK; hPAK65; Kinase; p21 (CDKN1A)-activated kinase 2a; p21 activated kinase 2; p21 protein (Cdc42/Rac)-activated kinase 2; p21 protein Cdc42 Rac activated kinase 2; p21-activated kinase, 65-KD; p21-activated protein kinase I; p21CDKN1A activated kinase 2; p58; p65PAK; PAK 2; PAK65; PAKgamma; S6 H4 kinase; Serine threonine protein kinase PAK 2; Serine/threonine-protein kinase PAK 3; p21-activated kinase 3; PAK-3; Beta-PAK; PAK1_HUMAN; PAK2_HUMAN.

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, Chicken, Dog, Pig, Cow, 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) 細(xì)胞凋亡 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 Thr423 

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1/2抗體費用 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 In mammals, there are several identified isoforms of p21 Activated Protein Kinases or PAKs: PAK1 and PAK3 are mostly brain specific, while PAK2 is expressed ubiquitously. Mutations of the gene coding for PAK3 are associated with X linked mental retardation and PAK3 is a key regulator of synapse formation and plasticity in the hippocampus. PAK3 is thought to play a key role in regulation of cell shape and motility as well as cell death. P21-activated kinase (PAK) is actually a family of serine/threonine protein kinases, members of which are activated by small molecular weight GTPases. The three most common isoforms are PAK 1, PAK 2, and PAK 3 (also known as alpha PAK, gamma PAK, and beta PAK, respectively). These kinases contain numerous regulatory elements that trigger diverse signaling processes such as those initiated by activated GTPases, interaction with Src homology 3 (SH3) domains, and caspase mediated proteolytic cleavage. Autophosphorylation of serine 141 (serine 144 for PAK 1 and serine 139 PAK 3), catalyzed by Cdc42, is required for activation of PAK.

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(xì)胞活動。包括細(xì)胞骨架的動力學(xué)調(diào)節(jié),細(xì)胞移動,生存和凋亡,細(xì)胞周期,基因轉(zhuǎn)錄調(diào)節(jié),細(xì)胞生長信號轉(zhuǎn)導(dǎo)和轉(zhuǎn)化等。

Anti-EGF/FITC 熒光素標(biāo)記表皮生長因子抗體(大鼠)IgGMulti-class antibodies規(guī)格： 0.2ml

Anti-HAVCR1/KIM1/TIM1/FITC 熒光素標(biāo)記病毒細(xì)胞受體1IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh C20orf194 20號染色體開放閱讀框194抗體 規(guī)格 0.2ml

Trk C 神經(jīng)生長因子受體的一種(抗原) 0.5mg

HSD3a 英文名稱： 羥基類固醇脫氫酶3α抗體 0.2ml

Rhesus antibody Rh S100A14 S100A14/15抗體 規(guī)格 0.1ml

Anti-HAVCR1/KIM1/TIM1/FITC 熒光素標(biāo)記病毒細(xì)胞受體1IgGMulti-class antibodies規(guī)格： 0.2ml

Anti-Phospho-Lyn (Tyr507) /FITC 熒光素標(biāo)記膜相關(guān)蛋白酪氨酸激酶Lyn抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Anti-phospho-Tau protein (Ser422)/FITC 熒光素標(biāo)記磷酸化微管相關(guān)蛋白抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh ADCK3/CABC1 伴侶蛋白bc1同源復(fù)合體抗體 規(guī)格 0.2ml

Rabbit Anti-dog IgG/FITC FITC標(biāo)記的兔抗狗IgG 0.3ml

GBP5 英文名稱： G蛋白結(jié)合蛋白5抗體 0.2ml

Rhesus antibody Rh Rabbit Anti-chicken IgG/Cy7 Cy7標(biāo)記的兔抗雞IgG 規(guī)格 0.1ml

Anti-phospho-Tau protein (Ser422)/FITC 熒光素標(biāo)記磷酸化微管相關(guān)蛋白抗體IgGMulti-class antibodies規(guī)格： 0.2ml

DUTP(Human deoxyuridinetriphate) ELISA Kit 人脫氧尿三磷酸Multi-class antibodies規(guī)格： 48T

Anti-PDGF-A 血小板源性生長因子-A抗體Multi-class antibodies規(guī)格： 0.1ml

Rhesus antibody Rh HMGB4 高遷移率族蛋白B4抗體 規(guī)格 0.1ml

AIRA(Human anti-insulin receptor antibody) ELISA Kit 人抗受體抗體 96T

phospho-PFKL (Ser775) 英文名稱： 磷酸化6磷酸果糖激酶抗體 0.1ml

C1orf31 英文名稱： 1號染色體開放閱讀框31抗體 0.2ml

Anti-PDGF-A 血小板源性生長因子-A抗體Multi-class antibodies規(guī)格： 0.1ml

CL-0316A7r5(大鼠胸大動脈平滑肌細(xì)胞)5×106cells/瓶×2

CD63 Others Rat 大鼠 CD63 / Tspan-30 / Teaspanin-30 人細(xì)胞裂解液 (陽性對照)

人海馬趾星形膠質(zhì)細(xì)胞總RNAHA-h NA

盤羊皮膚細(xì)胞;ASHS3 卵巢細(xì)胞，Lec1細(xì)胞 L7712細(xì)胞，615小鼠T細(xì)胞性瘤株

SV40T轉(zhuǎn)化的人胚腎細(xì)胞;293T

HA Others H5N1 甲型 H5N1 (A/Egypt/N05056/2009) 血凝素 (Hemagglutinin / HA) 人細(xì)胞裂解液 (陽性對照)

CL-03142V6.11(人胚腎細(xì)胞)5×106cells/瓶×2

HA Others H7N7 甲型 H7N7 (A/chicken/Netherlands/1/03) 血凝素 (Hemagglutinin / HA) 桿狀病毒-昆蟲細(xì)胞裂解液 (陽性對照)

人小腦星形膠質(zhì)細(xì)胞總RNAHA-c NA

J82細(xì)胞，細(xì)胞 人胚肺細(xì)胞,WI-38細(xì)胞 原代系膜細(xì)胞特制基礎(chǔ)無血清培養(yǎng)基Many types of cells包裝：500/100ml

豹貓肌肉成纖維樣細(xì)胞;LCM4

WISP1 Others Human 人 WISP1 / CCN4 人細(xì)胞裂解液 (陽性對照)

磷酸化p21激活激酶1/2抗體費用 SRC Others Human 人 SRC Kinase / Proto-oncogene c-Src 桿狀病毒-昆蟲細(xì)胞裂解液 (陽性對照)

Asp2人胚胎腎細(xì)胞轉(zhuǎn)化細(xì)胞;FC33

CM-H022人成纖維細(xì)胞完全培養(yǎng)基100mL

kasumi-1, 人細(xì)胞株 橫紋肌癌細(xì)胞,A673細(xì)胞 穩(wěn)定表達(dá)EBNA1的人胚腎細(xì)胞;293E

615小鼠株;Ca759

OSTM1 Others Human 人 OSTM1 人細(xì)胞裂解液 (陽性對照)

抗體的生物素化標(biāo)記實驗要點：

1. 磷酸化p21激活激酶1/2抗體費用 如在反應(yīng)混合液中有疊氮鈉或游離氨基存在,會抑制標(biāo)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(biāo)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(tuán),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(xì)胞的熒光標(biāo)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(xì)胞可導(dǎo)致高本底。

抗體的鑒定：

1）磷酸化p21激活激酶1/2抗體費用 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(kuò)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(kuò)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?？贵w的特異性高,它的識別能力就強(qiáng)。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?？贵w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。