抗体的生物素化标记实验要点：

1. 腺苷单磷酸活化蛋白激酶α2抗体说明书 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-PRKAA2/AMPK alpha 2 

中文名称   腺苷单磷酸活化蛋白激酶α2抗体说明书 

别    名  AMPK 2; AMPK2; AMPKalpha2; PRKAA 2; PRKAA; PRKAA2; Protein kinase AMP activated alpha 2 catalytic subunit; AAPK2_HUMAN.

浓    度   1mg/1ml

规 格  0.1ml/100μg 0.2ml/200μg

抗体来源   Rabbit

克隆类型   polyclonal

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

产品类型   一抗  

研究领域     细胞生物 神经生物学 信号转导 激酶和磷酸酶 Alzheimer's

蛋白分子量  predicted molecular weight: 62kDa

性    状   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human AMPK alpha 2 C-terminus 

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

腺苷单磷酸活化蛋白激酶α2抗体说明书 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 AMP activated protein kinase (AMPK) in mammals, and its homologue in Saccharomyces cerevisiae, are activated by cellular stresses associated with ATP depletion. AMPK is a heterotrimer comprising a catalytic alpha subunit with associated beta and gamma subunits. The alpha subunit has at least two isoforms (alpha 1 and alpha 2), which differ in their AMP dependence and subcellular localization, with alpha 2 complexes being partly nuclear. AMPK is activated allosterically by 5' AMP, which also promotes phosphorylation and activation by an upstream kinase, and inhibits dephosphorylation and inactivation. Elevation of AMP always accompanies depletion of ATP due to the action of adenylate kinase. Since high ATP antagonizes the activating effects of AMP, the system behaves like a cellular 'fuel gauge'. It is activated by various types of stress associated with ATP depletion, such as hypoxia, heat shock, metabolic poisoning and, in muscle, exercise. AMPK phosphorylates multiple targets which switch off anabolic pathways and switch on alternative catabolic pathways.

Function : Catalytic subunit of AMP-activated protein kinase (AMPK), an energy sensor protein kinase that plays a key role in regulating cellular energy metabolism. In response to reduction of intracellular ATP levels, AMPK activates energy-producing pathways and inhibits energy-consuming processes: inhibits protein, carbohydrate and lipid biosynthesis, as well as cell growth and proliferation. AMPK acts via direct phosphorylation of metabolic enzymes, and by longer-term effects via phosphorylation of transcription regulators. Also acts as a regulator of cellular polarity by remodeling the actin cytoskeleton; probably by indirectly activating myosin. Regulates lipid synthesis by phosphorylating and inactivating lipid metabolic enzymes such as ACACA, ACACB, GYS1, HMGCR and LIPE; regulates fatty acid and cholesterol synthesis by phosphorylating acetyl-CoA carboxylase (ACACA and ACACB) and hormone-sensitive lipase (LIPE) enzymes, respectively. Regulates insulin-signaling and glycolysis by phosphorylating IRS1, PFKFB2 and PFKFB3. AMPK stimulates glucose uptake in muscle by increasing the translocation of the glucose transporter SLC2A4/GLUT4 to the plasma membrane, possibly by mediating phosphorylation of TBC1D4/AS160. Regulates transcription and chromatin structure by phosphorylating transcription regulators involved in energy metabolism such as CRTC2/TORC2, FOXO3, histone H2B, HDAC5, MEF2C, MLXIPL/ChREBP, EP300, HNF4A, p53/TP53, SREBF1, SREBF2 and PPARGC1A. Acts as a key regulator of glucose homeostasis in liver by phosphorylating CRTC2/TORC2, leading to CRTC2/TORC2 sequestration in the cytoplasm. In response to stress, phosphorylates 'Ser-36' of histone H2B (H2BS36ph), leading to promote transcription. Acts as a key regulator of cell growth and proliferation by phosphorylating TSC2, RPTOR and ATG1: in response to nutrient limitation, negatively regulates the mTORC1 complex by phosphorylating RPTOR component of the mTORC1 complex and by phosphorylating and activating TSC2. In response to nutrient limitation, promotes autophagy by phosphorylating and activating ULK1. AMPK also acts as a regulator of circadian rhythm by mediating phosphorylation of CRY1, leading to destabilize it. May regulate the Wnt signaling pathway by phosphorylating CTNNB1, leading to stabilize it. Also phosphorylates CFTR, EEF2K, KLC1, NOS3 and SLC12A1.

Subunit : AMPK is a heterotrimer of an alpha catalytic subunit (PRKAA1 or PRKAA2), a beta (PRKAB1 or PRKAB2) and a gamma non-catalytic subunits (PRKAG1, PRKAG2 or PRKAG3). Interacts with FNIP1 and FNIP2.

Subcellular Location : Cytoplasm (By similarity). Nucleus. Note=In response to stress, recruited by p53/TP53 to specific promoters.

Phosphorylated at Thr-172 by STK11/LKB1 in complex with STE20-related adapter-alpha (STRADA) pseudo kinase and CAB39. Also phosphorylated at Thr-172 by CAMKK2; triggered by a rise in intracellular calcium ions, without detectable changes in the AMP/ATP ratio. CAMKK1 can also phosphorylate Thr-172, but at much lower lvel. Dephosphorylated by protein phosphatase 2A and 2C (PP2A and PP2C). Phosphorylated by ULK1; leading to negatively regulate AMPK activity and suggesting the existence of a regulatory feedback loop between ULK1 and AMPK.

Similarity : Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. SNF1 subfamily. [

Contains 1 protein kinase domain.

Database links : UniProtKB/Swiss-Prot: P54646.2

抗体的鉴定：

1）腺苷单磷酸活化蛋白激酶α2抗体说明书 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-BRCA2/FITC 荧光素标记癌易感基因2抗体IgGMulti-class antibodies规格： 0.2ml

Anti-GRM2/FITC 荧光素标记代谢型谷氨酸受体2IgGMulti-class antibodies规格： 0.2ml

Rhesus antibody Rh CIAPIN1 细胞因子诱导凋亡抑制因子1 规格 0.2ml

BKCa channels(calcium-activated potassium channel) 钙激活通道蛋白 0.5mg

IGFL4 英文名称： 生长因子样家族成员4抗体 0.2ml

Rhesus antibody Rh TNIK TRAF2和NCK激酶相互作用蛋白抗体 规格 0.2ml

Anti-GRM2/FITC 荧光素标记代谢型谷氨酸受体2IgGMulti-class antibodies规格： 0.2ml

Anti-ICAD/FITC 荧光素标记Caspase 激活的脱氧核糖核酸酶抑制剂抗体IgGMulti-class antibodies规格： 0.2ml

Anti-Phospho-PRAS40 (Thr246)/FITC 荧光素标记磷酸化蛋白激酶AKT底物1抗体IgGMulti-class antibodies规格： 0.2ml

Rhesus antibody Rh ARTS1 脂肪细胞源性亮氨酸氨基肽酶抗体(内皮细胞生长因子诱导蛋白) 规格 0.2ml

Donkey Anti-rabbit IgG/Cy7 Cy7标记的驴抗兔IgG 0.1ml

Gcn1l1 英文名称： 广泛控制基酸合成1样蛋白1抗体 0.2ml

Rhesus antibody Rh Rabbit anti-Monkey IgM whole serum 兔抗猴IgM抗血清 规格 1ml

Anti-Phospho-PRAS40 (Thr246)/FITC 荧光素标记磷酸化蛋白激酶AKT底物1抗体IgGMulti-class antibodies规格： 0.2ml

17-OHCS ELISA Kit 大鼠17羟皮质类固醇Multi-class antibodies规格： 48T

Anti-Agrin/FITC 荧光素标记聚集蛋白抗体IgGMulti-class antibodies规格： 0.2ml

Rhesus antibody Rh FBXO28 FBXO28蛋白抗体 规格 0.2ml

F III ELISA Kit 大鼠凝血因子Ⅲ 96T

phospho-MLF1 Interacting Protein(Thr78) 英文名称： 磷酸化卡波西氏病毒潜伏核抗原相互作用蛋白1抗体 0.1ml

AGRP 英文名称： 褐黑素相关蛋白ART抗体 0.2ml

Anti-Agrin/FITC 荧光素标记聚集蛋白抗体IgGMulti-class antibodies规格： 0.2ml

786-O [786-0], 人肾透明腺癌细胞

CD300LG Others Human 人 CLM-9 / EM4 / CD300LG 人细胞裂解液 (阳性对照)

表达SV40T和EBNA1的人胚肾细胞;293ET

KC细胞，人细胞 小鼠脑神经瘤细胞,Neuro-2a[N2a;Neuro-2a]细胞 CL-0373KiMA(人胚肾上皮细胞系)5×106cells/瓶×2

BGC-803(人细胞) 5×106cells/瓶×2

HWP-c subcutaneous 人类白前脂肪细胞(HWP)皮下 500,000cells 人淋巴成纤维细胞HLF

786-O [786-0], 人肾透明腺癌细胞

CD300LG Others Human 人 CLM-9 / EM4 / CD300LG 人细胞裂解液 (阳性对照)

表达SV40T和EBNA1的人胚肾细胞;293ET

KC细胞，人细胞 小鼠脑神经瘤细胞,Neuro-2a[N2a;Neuro-2a]细胞 CL-0373KiMA(人胚肾上皮细胞系)5×106cells/瓶×2

BGC-803(人细胞) 5×106cells/瓶×2

HWP-c subcutaneous 人类白前脂肪细胞(HWP)皮下 500,000cells 人淋巴成纤维细胞HLF

腺苷单磷酸活化蛋白激酶α2抗体说明书 TNFRSF13C Others Rat 大鼠 BAFFR / TNFRSF13C 人细胞裂解液 (阳性对照)

人食道上皮细胞总RNAHEEpiC NA

叙利亚仓鼠肾细胞;BHK-21

PC-2细胞，人*癌细胞 人细胞系,HS-4细胞 小鼠瘤株;B16

Hut-102(人皮肤T细胞) 5×106cells/瓶×2

Promocell C-22020 Endothelial Cell GrowthMedium MV, 内皮细胞生长培养基MV(即用型) 500ml