抗体的生物素化标记实验要点：

1. 磷酸化p21激活激酶1/2/3抗体品牌 如在反应混合液中有叠氮钠或游离氨基存在,会抑制标记反应。因此,蛋白质在反应前要对 0.1mol/L碳酸氢钠缓冲液或0.5mol/L硼酸缓冲液充分透析；

2.所用的NHSB及待生物素化蛋白质之间的分子比按蛋白质表面的ε-氨基的密度会有所不同,选择不当则影响标记的效率,应先用几个不同的分子比来筛选最适条件；

3.用NHSB量过量也是不利的,抗原的结合位点可能因此被封闭,导致抗体失活；

4.由于抗体的氨基不易接近可能造成生物素化不足,此时可加入去污剂如 Triton x-100, Tween20等；

5.当游离ε-氨基(赖氨酸残基的氨基)存在于抗体的抗原结合位点时,或位于酶的催化位点时,生物素化会降低或损伤抗体蛋白的结合力或活性；

6.生物素还可能与不同的功能基团,如羰基、氨基、巯基、异咪唑基及*基,也可与糖基共价结合；

7.交联反应后,应充分透析,否则,残余的生物素会对生物素化抗体与亲和素的结合产生竞争作用；

8.在细胞的荧光标记实验中,中和亲和素的本底低,但由于链霉亲和素含有少量正电荷,故对某些细胞可导致高本底。

产品订购信息：
英文名称  Anti-phospho-PAK1/2/3(Thr423) 

中文名称   磷酸化p21激活激酶1/2/3抗体品牌 

别    名  phospho-PAK1(pSer144)+Anti-phospho-PAK2(pSer141)+Anti-phospho-PAK3(pSer139); Alpha PAK; Beta PAK; Gamma PAK; Oligophrenin 3; OPHN3; p21 activated kinase 1; p21 Activated Kinase 1 + 2 + 3; p21 activated kinase 2 ; p21 activated kinase 3; P65 PAK; PAK1; PAK2; PAK3; PAK3beta; PAK65; PAKalpha; PAKgamma; S6/H4 kinase; Serine/threonine protein kinase PAK 1; Serine/threonine protein kinase PAK 2; Serine/threonine protein kinase PAK 3; Serine/threonine protein kinase PAK1; Serine/threonine protein kinase PAK2; Serine/threonine protein kinase PAK3; PAK1_HUMAN; PAK2_HUMAN; PAK3_HUMAN.

浓    度   1mg/1ml

规 格  0.1ml/100μg

抗体来源   Rabbit

克隆类型   polyclonal

交叉反应   Human, Mouse, Rat, Chicken, Dog, Pig, Cow, Rabbit 

产品类型   一抗 磷酸化抗体  

研究领域     细胞生物 神经生物学 信号转导 细胞凋亡 转录调节因子 激酶和磷酸酶

蛋白分子量  predicted molecular weight: 60kDa

性    状   Lyophilized or Liquid

免 疫 原 KLH conjugated Synthesised phosphopeptide derived from human PAK1 around the phosphorylation site of (Thr423) 

亚    型   IgG

纯化方法   affinity purified by Protein A

储 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1/2/3抗体品牌 产品应用    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

（石蜡切片需做抗原修复） 

 not yet tested in other applications.

 optimal dilutions/concentrations should be determined by the end user.  

保存条件  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. 

产品介绍 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.

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.

Post-translational modifications : Autophosphorylated when activated by CDC42/p21 and RAC1. Phosphorylation at Thr-423 by PDPK1 results in its 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蛋白具有广泛的生物学功能,也是一个保守的丝氨酸/苏氨酸蛋白激酶,参与许多重要的细胞活动。包括细胞骨架的动力学调节,细胞移动,生存和凋亡,细胞周期,基因转录调节,细胞生长信号转导和转化等。

抗体的鉴定：

1）磷酸化p21激活激酶1/2/3抗体品牌 抗体的效价鉴定：不管是用于诊断还是用于,制备抗体的目的都是要求较高效价。不同的抗原制备的抗体,要求的效价不一。鉴定效价的方法很多,包括有试管凝集反应,琼脂扩散试验,酶联免疫吸附试验等。常用的抗原所制备的抗体一般都有约成的鉴定效价的方法,以资比较。如制备抗抗体的效价,一般就采用琼脂扩散试验来鉴定。

2）抗体的特异性鉴定：抗体的特异性是指与相应抗原或近似抗原物质的识别能力。抗体的特异性高,它的识别能力就强。衡量特异性通常以交叉反应率来表示。交叉反应率可用竞争抑制试验测定。以不同浓度抗原和近似抗原分别做竞争抑制曲线,计算各自的结合率,求出各自在IC50时的浓度,并按公式计算交叉反应率。 

如果所用抗原浓度IC50浓度为pg/管,而一些近似抗原物质的IC50浓度几乎是无穷大时,表示这一抗血清与其他抗原物质的交叉反应率近似为0,即该血清的特异性较好。

3）抗体亲和力：是指抗体和抗原结合的牢固程度。亲和力的高低是由抗原分子的大小,抗体分子的结合位点与抗原决定簇之间立体构型的合适度决定的。有助于维持抗原抗体复合物稳定的分子间力有氢键,疏水键,侧链相反电荷基因的库仑力,范德华力和空间斥力。亲和力常以亲和常数K表示,K的单位是L/mol。抗体亲和力的测定对抗体的筛选,确定抗体的用途,验证抗体的均一性等均有重要意义。

Anti-Phospho-C/EBPalpha (Ser21) /FITC 荧光素标记兔抗人、大、小鼠磷酸化转录调节因子C/EBP α 抗体IgGMulti-class antibodies规格： 0.2ml

Anti-IL-17C/FITC 荧光素标记白介素-17C抗体IgGMulti-class antibodies规格： 0.2ml

Rhesus antibody Rh CD158e NK细胞抑制性受体3DL1抗体 规格 0.2ml

LN (laminin) 层粘连蛋白(多肽抗原) 0.5mg

IGF2BP1 英文名称： 样生长因子ⅡmRNA结合蛋白1抗体 0.2ml

Rhesus antibody Rh SSEA3 阶段特异性胚胎表面抗原-3抗体 规格 0.1ml

Anti-IL-17C/FITC 荧光素标记白介素-17C抗体IgGMulti-class antibodies规格： 0.2ml

Anti-IL-17(Interleukin-17)human/FITC 荧光素标记白介素-17抗体(人)IgGMulti-class antibodies规格： 0.2ml

Anti-Phospho-Rad17 (Ser645) /FITC 荧光素标记磷酸化细胞周期检控Rad17蛋白抗体IgGMulti-class antibodies规格： 0.2ml

Rhesus antibody Rh APE/Girdin 肌动蛋白结合蛋白Girdin抗体 规格 0.1ml

Rabbit Anti-rat IgG/Alexa Fluor 647 Alexa Fluor 647标记的兔抗大鼠IgG 0.1ml

GIMAP7 英文名称： GTP酶IMAP家族成员7抗体 0.2ml

Rhesus antibody Rh Rabbit Anti-human IgM/PE-Cy3 PE-Cy3标记的兔抗人IgM 规格 0.1ml

Anti-Phospho-Rad17 (Ser645) /FITC 荧光素标记磷酸化细胞周期检控Rad17蛋白抗体IgGMulti-class antibodies规格： 0.2ml

TrxR(Mouse thioredoxin reductase) ELISA Kit 小鼠硫氧还蛋白还原酶Multi-class antibodies规格： 48T

Anti-CBL2 /FITC 荧光素标记兔抗人、大、小鼠原癌蛋白CBL2抗体IgGMulti-class antibodies规格： 0.2ml

Rhesus antibody Rh Egr1 早期生长应答蛋白1抗体 规格 0.1ml

CR2/sCD21(Human Soluble Cluster of differentiation 21) ELISA Kit 人可溶性CD21 96T

Phospho-MYL(Ser19) 英文名称： 磷酸化肌球蛋白轻链2抗体 0.1ml

AF10 英文名称： 髓系、淋巴、混合系易位基因10抗体 0.1ml

Anti-CBL2 /FITC 荧光素标记兔抗人、大、小鼠原癌蛋白CBL2抗体IgGMulti-class antibodies规格： 0.2ml

C6, 大鼠脑细胞

FCGRT & B2M Others Mouse 小鼠 FCGRT & B2M Heterodimer 人细胞裂解液 (阳性对照)

大鼠脑内皮细胞完全培养基 100mL

LLC-PK1猪肾近曲小管上皮细胞 LLC-PK1 porcine kidney proximal tubular epithelial cells M199+3% FBS/DMEM+F12+12%FBS

IFNB1 Protein Mouse 重组小鼠 IFNB1 / IFN-beta / Ierferon beta 蛋白 (Fc 标签)

MDA-MB-468-06(人癌细胞) 5×106cells/瓶×2 Y1(肾上腺皮质细胞)

C57小鼠瘤株;ME (Me)

OLR1 Others Human 人 OLR1 / LOX1 人细胞裂解液 (阳性对照)

CaEs-17 人细胞

GBC-SD人细胞 GBC-SD cells in human gallbladder carcinomas RPMI-1640(GIBCO)+10%FBS

LEFTY2 Protein Human 重组人 Lefty2 / Lefty A 蛋白 (Fc 标签)

人肝内胆管上皮细胞 (HIBEC)( 5×105 ) HSC-T6, 大鼠肝星状细胞系 Rattus

磷酸化p21激活激酶1/2/3抗体品牌 MAPKAPK5 Others Human 人 MAPKAPK5 杆状病毒-昆虫细胞裂解液 (阳性对照)

叙利亚仓鼠肌肉细胞;GH-M1

CL-0081F81(猫肾细胞)5×106cells/瓶×2

KM3, 人细胞 细胞,HCC-9204细胞 NCI-H292(细胞(淋巴结转移))

小鼠细胞;GT1-1

CD2 Others Human 人 CD2 人细胞裂解液 (阳性对照)