抗體的生物素化標(biāo)記實(shí)驗(yàn)要點(diǎn)：

1. 腺苷單磷酸活化蛋白激酶α2抗體說(shuō)明書(shū) 如在反應(yīng)混合液中有疊氮鈉或游離氨基存在,會(huì)抑制標(biāo)記反應(yīng)。因此,蛋白質(zhì)在反應(yīng)前要對(duì) 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(huì)有所不同,選擇不當(dāng)則影響標(biāo)記的效率,應(yīng)先用幾個(gè)不同的分子比來(lái)篩選最適條件；

3.用NHSB量過(guò)量也是不利的,抗原的結(jié)合位點(diǎn)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(shí)可加入去污劑如 Triton x-100, Tween20等；

5.當(dāng)游離ε-氨基(賴(lài)氨酸殘基的氨基)存在于抗體的抗原結(jié)合位點(diǎn)時(shí),或位于酶的催化位點(diǎn)時(shí),生物素化會(huì)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(jià)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(huì)對(duì)生物素化抗體與親和素的結(jié)合產(chǎn)生競(jìng)爭(zhēng)作用；

8.在細(xì)胞的熒光標(biāo)記實(shí)驗(yàn)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(duì)某些細(xì)胞可導(dǎo)致高本底。

產(chǎn)品訂購(gòu)信息：
英文名稱(chēng)  Anti-PRKAA2/AMPK alpha 2 

中文名稱(chēng)   腺苷單磷酸活化蛋白激酶α2抗體說(shuō)明書(shū) 

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

濃    度   1mg/1ml

規(guī) 格  0.1ml/100μg 0.2ml/200μg

抗體來(lái)源   Rabbit

克隆類(lèi)型   polyclonal

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

產(chǎn)品類(lèi)型   一抗  

研究領(lǐng)域     細(xì)胞生物 神經(jīng)生物學(xué) 信號(hào)轉(zhuǎn)導(dǎo) 激酶和磷酸酶 Alzheimer's

蛋白分子量  predicted molecular weight: 62kDa

性    狀   Lyophilized or Liquid

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

亞    型   IgG

純化方法   affinity purified by Protein A

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

腺苷單磷酸活化蛋白激酶α2抗體說(shuō)明書(shū) 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 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

抗體的鑒定：

1）腺苷單磷酸活化蛋白激酶α2抗體說(shuō)明書(shū) 抗體的效價(jià)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(jià)。不同的抗原制備的抗體,要求的效價(jià)不一。鑒定效價(jià)的方法很多,包括有試管凝集反應(yīng),瓊脂擴(kuò)散試驗(yàn),酶聯(lián)免疫吸附試驗(yàn)等。常用的抗原所制備的抗體一般都有約成的鑒定效價(jià)的方法,以資比較。如制備抗抗體的效價(jià),一般就采用瓊脂擴(kuò)散試驗(yàn)來(lái)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(shí)別能力?？贵w的特異性高,它的識(shí)別能力就強(qiáng)。衡量特異性通常以交叉反應(yīng)率來(lái)表示。交叉反應(yīng)率可用競(jìng)爭(zhēng)抑制試驗(yàn)測(cè)定。以不同濃度抗原和近似抗原分別做競(jìng)爭(zhēng)抑制曲線(xiàn),計(jì)算各自的結(jié)合率,求出各自在IC50時(shí)的濃度,并按公式計(jì)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(wú)窮大時(shí),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(diǎn)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(kù)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(cè)定對(duì)抗體的篩選,確定抗體的用途,驗(yàn)證抗體的均一性等均有重要意義。

Anti-BRCA2/FITC 熒光素標(biāo)記癌易感基因2抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Anti-GRM2/FITC 熒光素標(biāo)記代謝型谷氨酸受體2IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh CIAPIN1 細(xì)胞因子誘導(dǎo)凋亡抑制因子1 規(guī)格 0.2ml

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

IGFL4 英文名稱(chēng)： 生長(zhǎng)因子樣家族成員4抗體 0.2ml

Rhesus antibody Rh TNIK TRAF2和NCK激酶相互作用蛋白抗體 規(guī)格 0.2ml

Anti-GRM2/FITC 熒光素標(biāo)記代謝型谷氨酸受體2IgGMulti-class antibodies規(guī)格： 0.2ml

Anti-ICAD/FITC 熒光素標(biāo)記Caspase 激活的脫氧核糖核酸酶抑制劑抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Anti-Phospho-PRAS40 (Thr246)/FITC 熒光素標(biāo)記磷酸化蛋白激酶AKT底物1抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh ARTS1 脂肪細(xì)胞源性亮氨酸氨基肽酶抗體(內(nèi)皮細(xì)胞生長(zhǎng)因子誘導(dǎo)蛋白) 規(guī)格 0.2ml

Donkey Anti-rabbit IgG/Cy7 Cy7標(biāo)記的驢抗兔IgG 0.1ml

Gcn1l1 英文名稱(chēng)： 廣泛控制基酸合成1樣蛋白1抗體 0.2ml

Rhesus antibody Rh Rabbit anti-Monkey IgM whole serum 兔抗猴IgM抗血清 規(guī)格 1ml

Anti-Phospho-PRAS40 (Thr246)/FITC 熒光素標(biāo)記磷酸化蛋白激酶AKT底物1抗體IgGMulti-class antibodies規(guī)格： 0.2ml

17-OHCS ELISA Kit 大鼠17羥皮質(zhì)類(lèi)固醇Multi-class antibodies規(guī)格： 48T

Anti-Agrin/FITC 熒光素標(biāo)記聚集蛋白抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh FBXO28 FBXO28蛋白抗體 規(guī)格 0.2ml

F III ELISA Kit 大鼠凝血因子Ⅲ 96T

phospho-MLF1 Interacting Protein(Thr78) 英文名稱(chēng)： 磷酸化卡波西氏病毒潛伏核抗原相互作用蛋白1抗體 0.1ml

AGRP 英文名稱(chēng)： 褐黑素相關(guān)蛋白ART抗體 0.2ml

Anti-Agrin/FITC 熒光素標(biāo)記聚集蛋白抗體IgGMulti-class antibodies規(guī)格： 0.2ml

786-O [786-0], 人腎透明腺癌細(xì)胞

CD300LG Others Human 人 CLM-9 / EM4 / CD300LG 人細(xì)胞裂解液 (陽(yáng)性對(duì)照)

表達(dá)SV40T和EBNA1的人胚腎細(xì)胞;293ET

KC細(xì)胞，人細(xì)胞 小鼠腦神經(jīng)瘤細(xì)胞,Neuro-2a[N2a;Neuro-2a]細(xì)胞 CL-0373KiMA(人胚腎上皮細(xì)胞系)5×106cells/瓶×2

BGC-803(人細(xì)胞) 5×106cells/瓶×2

HWP-c subcutaneous 人類(lèi)白前脂肪細(xì)胞(HWP)皮下 500,000cells 人淋巴成纖維細(xì)胞HLF

786-O [786-0], 人腎透明腺癌細(xì)胞

CD300LG Others Human 人 CLM-9 / EM4 / CD300LG 人細(xì)胞裂解液 (陽(yáng)性對(duì)照)

表達(dá)SV40T和EBNA1的人胚腎細(xì)胞;293ET

KC細(xì)胞，人細(xì)胞 小鼠腦神經(jīng)瘤細(xì)胞,Neuro-2a[N2a;Neuro-2a]細(xì)胞 CL-0373KiMA(人胚腎上皮細(xì)胞系)5×106cells/瓶×2

BGC-803(人細(xì)胞) 5×106cells/瓶×2

HWP-c subcutaneous 人類(lèi)白前脂肪細(xì)胞(HWP)皮下 500,000cells 人淋巴成纖維細(xì)胞HLF

腺苷單磷酸活化蛋白激酶α2抗體說(shuō)明書(shū) TNFRSF13C Others Rat 大鼠 BAFFR / TNFRSF13C 人細(xì)胞裂解液 (陽(yáng)性對(duì)照)

人食道上皮細(xì)胞總RNAHEEpiC NA

敘利亞倉(cāng)鼠腎細(xì)胞;BHK-21

PC-2細(xì)胞，人*癌細(xì)胞 人細(xì)胞系,HS-4細(xì)胞 小鼠瘤株;B16

Hut-102(人皮膚T細(xì)胞) 5×106cells/瓶×2

Promocell C-22020 Endothelial Cell GrowthMedium MV, 內(nèi)皮細(xì)胞生長(zhǎng)培養(yǎng)基MV(即用型) 500ml