Anti-phospho-AKT1 (Tyr474)/FIT

产品名称	Anti-phospho-AKT1 (Tyr474)/FITC抗体
英文名称	Anti-phospho-AKT1 (Tyr474)/FITC
货号	XGK3485

英文名称  Anti-phospho-AKT1 (Tyr474)/FITC
中文名称  FITC标记的磷酸化蛋白激酶AKT1抗体 
别    名AKT1 (phospho Y474); AKT1(phospho-Tyr474); AKT3 (phospho Y473); p-AKT3 (phospho Y473); phospho-AKT1(Tyr474); phospho-AKT2 (Tyr475); AKT1(phospho Tyr474); AKT2 (phospho Tyr475); AKT3(phospho Tyr473); AKT 1; AKT; AKT1; AKT-1; AKT1_HUMAN; C AKT; cAKT; MGC9965; MGC99656; Oncogene AKT1; PKB; PKB alpha; PKB-ALPHA; PRKBA; Protein Kinase B Alpha; Protein kinase B; Proto-oncogene c-Akt; RAC Alpha; RAC alpha serine/threonine protein kinase; RAC; RAC PK Alpha; Rac protein kinase alpha; RAC Serine/Threonine Protein Kinase; RAC-alpha serine/threonine-protein kinase; RAC-PK-alpha; v akt murine thymoma viral oncogene homolog 1; vAKT Murine Thymoma Viral Oncogene Homolog 1.
规格:100ul
说 明 书100ul
产品类型磷酸化抗体
研究领域肿瘤  细胞生物  信号转导  细胞凋亡  转录调节因子
抗体来源Rabbit
克隆类型Polyclonal
交叉反应 Human, Mouse, Rat, Chicken, Dog, Pig, Cow, Rabbit, Sheep,
产品应用Flow-Cyt=1:50-200 ICC=1:50-200 IF=1:50-200
not yet tested in other applications.
optimal dilutions/concentrations should be determined by the end user.
分 子 量56kDa
细胞定位细胞膜
性    状Lyophilized or Liquid
浓    度1mg/ml
免 疫 原KLH conjugated synthesised phosphopeptide derived from human AKT1 around the phosphorylation site of Tyr474
亚    型IgG
纯化方法affinity purified by Protein A
储 存 液0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol.
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.
产品介绍background:
AKT, also known as protein kinase B (PKB), is a 57 kDa serine/threonine protein kinase. There are three mammalian isoforms of Akt: AKT1 (PKB alpha), AKT2 (PKB beta) and AKT3 (PKB gamma) with AKT2 and AKT3 being approximately 82% identical with the AKT1 isoform. Each isoform has a pleckstrin homology (PH)domain, a kinase domain and a carboxy terminal regulatory domain. AKT was originally cloned from the retrovirus AKT8, and is a key regulator of many signal transduction pathways. Its tight control over cell proliferation and cell viability are manifold; overexpression or inappropriate activation of AKT has been seen in many types of cancer. AKT mediates many of the downstream events of phosphatidylinositol 3 kinase (a lipid kinase activated by growth factors, cytokines and insulin). PI3 kinase recruits AKT to the membrane, where it is activated by PDK1 phosphorylation. Once phosphorylated, AKT dissociates from the membrane and phosphorylates targets in the cytoplasm and the cell nucleus
Function:
AKT1 is one of 3 closely related serine/threonine-protein kinases (AKT1, AKT2 and AKT3) called the AKT kinase, and which regulate many processes including metabolism, proliferation, cell survival, growth and angiogenesis. This is mediated through serine and/or threonine phosphorylation of a range of downstream substrates. Over 100 substrate candidates have been reported so far, but for most of them, no isoform specificity has been reported. AKT is responsible of the regulation of glucose uptake by mediating insulin-induced translocation of the SLC2A4/GLUT4 glucose transporter to the cell surface. Phosphorylation of PTPN1 at 'Ser-50' negatively modulates its phosphatase activity preventing dephosphorylation of the insulin receptor and the attenuation of insulin signaling. Phosphorylation of TBC1D4 triggers the binding of this effector to inhibitory 14-3-3 proteins, which is required for insulin-stimulated glucose transport. AKT regulates also the storage of glucose in the form of glycogen by phosphorylating GSK3A at 'Ser-21' and GSK3B at 'Ser-9', resulting in inhibition of its kinase activity. Phosphorylation of GSK3 isoforms by AKT is also thought to be one mechanism by which cell proliferation is driven. AKT regulates also cell survival via the phosphorylation of MAP3K5 (apoptosis signal-related kinase). Phosphorylation of 'Ser-83' decreases MAP3K5 kinase activity stimulated by oxidative stress and thereby prevents apoptosis. AKT mediates insulin-stimulated protein synthesis by phosphorylating TSC2 at 'Ser-939' and 'Thr-1462', thereby activating mTORC1 signaling and leading to both phosphorylation of 4E-BP1 and in activation of RPS6KB1. AKT is involved in the phosphorylation of members of the FOXO factors (Forkhead family of transcription factors), leading to binding of 14-3-3 proteins and cytoplasmic localization. In particular, FOXO1 is phosphorylated at 'Thr-24', 'Ser-256' and 'Ser-319'. FOXO3 and FOXO4 are phosphorylated on equivalent sites. AKT has an important role in the regulation of NF-kappa-B-dependent gene transcription and positively regulates the activity of CREB1 (cyclic AMP (cAMP)-response element binding protein). The phosphorylation of CREB1 induces the binding of accessory proteins that are necessary for the transcription of pro-survival genes such as BCL2 and MCL1. AKT phosphorylates 'Ser-454' on ATP citrate lyase (ACLY), thereby potentially regulating ACLY activity and fatty acid synthesis. Activates the 3B isoform of cyclic nucleotide phosphodiesterase (PDE3B) via phosphorylation of 'Ser-273', resulting in reduced cyclic AMP levels and inhibition of lipolysis. Phosphorylates PIKFYVE on 'Ser-318', which results in increased PI(3)P-5 activity. The Rho GTPase-activating protein DLC1 is another substrate and its phosphorylation is implicated in the regulation cell proliferation and cell growth. AKT plays a role as key modulator of the AKT-mTOR signaling pathway controlling the tempo of the process of newborn neurons integration during adult neurogenesis, including correct neuron positioning, dendritic development and synapse formation. Signals downstream of phosphatidylinositol 3-kinase (PI(3)K) to mediate the effects of various growth factors such as platelet-derived growth factor (PDGF), epidermal growth factor (EGF), insulin and insulin-like growth factor I (IGF-I). AKT mediates the antiapoptotic effects of IGF-I. Essential for the SPATA13-mediated regulation of cell migration and adhesion assembly and disassembly. May be involved in the regulation of the placental development. Phosphorylates STK4/MST1 at 'Thr-120' and 'Thr-387' leading to inhibition of its: kinase activity, nuclear translocation, autophosphorylation and ability to phosphorylate FOXO3. Phosphorylates STK3/MST2 at 'Thr-117' and 'Thr-384' leading to inhibition of its: cleavage, kinase activity, autophosphorylation at Thr-180, binding to RASSF1 and nuclear translocation. Phosphorylates SRPK2 and enhances its kinase activity towards SRSF2 and ACIN1 and promotes its nuclear translocation. Phosphorylates RAF1 at 'Ser-259' and negatively regulates its activity. Phosphorylation of BAD stimulates its pro-apoptotic activity.

实验原理 :
（1）特异性结合抗原：抗体本身不能直接溶解或杀伤带有特异抗原的靶细胞，通常需要补体或吞噬细胞等共同发挥效应以清除病原微生物或导致病理损伤。然而，抗体可通过与病毒或毒素的特异性结合，直接发挥中和病毒的作用。
（2）活补体：IgM、IgG1、IgG2和IgG3可通过经典途径激活补体，凝聚的IgA、IgG4和IgE可通过替代途径激活补体。
（3）结合细胞：不同类别的免疫球蛋白，可结合不同种的细胞，参与免疫应答。
（4）可通过胎盘及粘膜：免疫球蛋白G（IgG）能通过胎盘进入胎儿血流中，使胎儿形成自然被动
免疫。免疫球蛋白A（IgA）可通过消化道及呼吸道粘膜，是粘膜局部抗感染免疫的主要因素。
（5）具有抗原性：抗体分子是一种蛋白质，也具有刺机体产生免疫应答的性能。不同的免疫球蛋白分子，各具有不同的抗原性。
（6）抗体对理化因子的抵抗力与一般球蛋白相同：不耐热，60～70℃即被破坏。各种酶及能使蛋白质凝固变性的物质，均能破坏抗体的作用。抗体可被中性盐类沉淀。在上常可用硫酸铵或硫酸钠从免疫血清中沉淀出含有抗体的球蛋白，再经透析法将其纯化。
抗体制备过程：
1.材料与试剂
  a．提取的动物 Ig
  b．弗氏完全佐剂和弗氏不完全佐剂
  c．青霉素和链霉素
  d．实验动物 兔
  e．其它材料及试剂
2、选择实验活体。
3、进行动物免疫实验。
4、试取血样进行测试，查看免疫效果。
5、如果免疫成功，杀死实验活体，采集全部血清。
6、纯化出抗体。
7、鉴定抗体。胎牛血清（无菌采制）
我司提供以下技术外包服务:
1. Anti-phospho-AKT1 (Tyr474)/FITC抗体分子生物学：质粒抽提、PCR、Q-PCR、RT-PCR、分子生物学：基因合成、引物合成、基因测序、载体构建等
2.蛋白工程：原核、哺乳动物蛋白表达系统等
3.病毒包装：腺病毒、慢病毒等
4.抗体工程：磁珠分选、病理染色、WB、ELISA、IP、IF、IHC、FACS、Confocal等等
5.细胞工程：细胞表型分析(凋亡、增殖、周期、迁移、侵袭、修复、克隆形成)、细胞培养、细胞膜制备、稳定细胞株构建、细胞RNAi技术等等。
抗体的鉴定：
1）抗体的效价鉴定：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。
3）真核翻译起始因子2C2抗体的亲和力：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。
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Rat iggering receptor expressed on myeloid cells -1 (EM-1) ELISA Kit 大鼠髓系细胞触发受体-1(EM-1)ELISA试剂盒
HumanChorionicGonadoophin,HCGELISAKit 人绒毛膜促性腺激素(HCG)ELISA试剂盒 96T/48T 进口分装
HumanClusterofdiffereiation30,CD30ELISA试剂盒人CD30分子(CD30)ELISA试剂盒规格：96T/48T
组蛋白脱乙酰化酶10(HDAC10)活性比色法定量检测试剂盒20次
humahofamilyGTPase1,RND1ELISAKit人Rho家族GTP酶1(RND1)ELISA试剂盒规格：96T/48T
人谷酸脱羧酶自身抗体(GAD-Ab)ELISA 试剂盒 96T/48T 试剂盒 组装/原装
小鼠白介素29(IL-29)免疫试剂盒 Mouse Ierleukin 29,IL-29 ELISA Kit
心肌营养素1(CT-1)ELISA试剂盒 ，英文名： CT-1 ELISA Kit
Mouseinducibleniicoxidesyhase,iNOSELISA试剂盒小鼠诱导型合成酶(iNOS)ELISA试剂盒规格：96T/48T
ELISAKitBNP兔子脑素/脑尿肽规格：48T/96T
ChickenImmuneRNA,IrnaELISAKit鸡免疫核糖核酸(Irna)ELISA试剂盒规格：96T/48T
人可溶性白细胞分化抗原40配体(sCD40L)ELISA 试剂盒 96T/48T 试剂盒 组装/原装
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坏死因子α(TNF-α)ELISA试剂盒 ，英文名： TNF-α ELISA Kit
humanUDP-glucoseceramideglucosylansferaseELISA试剂盒人尿苷二0酸葡萄糖神经酰胺葡萄糖基转移酶(UGCG)ELISA试剂盒规格：96T/48T
ELISAKitPCNA小鼠抗增殖细胞核抗原抗体规格：48T/96T
Humanai-gasicparietalcellaibody,AGPA/PCAELISAKit人抗胃壁细胞抗体(AGPA/PCA)ELISA试剂盒规格：96T/48T
中国仓鼠卵巢细胞K1(亚系克隆);CHO-K1
ULBP2 Others Human 人 ULBP2 / N2DL-2 人细胞裂解液 (阳性对照)
Ke-3 人肾癌细胞
MDA-MB-436人腺癌细胞 MDA-MB-436 human breast adenocarcinoma cells L-15培养基+10%FBS
FAM3C Protein Human 重组人 FAM3C / ILEI 蛋白 (Fc 标签)
小鼠雪旺细胞(MSC)(5×105) MDA-MB-231, 人癌细胞 Human
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人脐静脉平滑肌细胞
人毛发干细胞(HHDPC)( 5×105 )
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CDH17 Others Human 人 CDH17 / Cadherin-17 / LI-cadherin 人细胞裂解液 (阳性对照)