抗体的生物素化标记实验要点：

1. 磷酸化核糖体S6蛋白激酶β2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-Phospho-p70 S6 Kinase Beta (Thr444 + Ser447) 

中文名称   磷酸化核糖体S6蛋白激酶β2抗体说明书 

别    名  p70 S6 Kinase(Phospho-Thr444 + Ser447); P70 S6 Kinase beta (phospho T444 + S447); 70 kDa ribosomal protein S6 kinase 2; p70 beta; p70 ribosomal S6 kinase beta; p70 S6 kinase beta; p70 S6KB; p70 S6Kbeta; p70(S6K) beta; p70-beta; p70-S6KB; p70S6Kb; Ribosomal protein S6 kinase 70kDa, polypeptide 2; Ribosomal protein S6 kinase beta 2; Ribosomal protein S6 kinase beta-2; RPS6KB2; S6 kinase related kinase; S6 kinase-related kinase; S6K beta 2; S6K beta; S6K-beta; S6K2; Serine/threonine protein kinase 14 beta; Serine/threonine-protein kinase 14 beta; SRK; STK14B; KS6B1_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, Horse, Rabbit 

产品类型   一抗 磷酸化抗体  

研究领域     免疫学 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: 70kDa

性    状   Lyophilized or Liquid

免 疫 原 KLH conjugated Synthesised phosphopeptide derived from human p70 S6 Kinase Beta around the phosphorylation site of Thr444/Ser447 

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

磷酸化核糖体S6蛋白激酶β2抗体说明书 产品应用    WB=1:100-500 ELISA=1:500-1000 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 This gene encodes a member of the ribosomal S6 kinase family of serine/threonine kinases. The encoded protein responds to mTOR (mammalian target of rapamycin) signaling to promote protein synthesis, cell growth, and cell proliferation. Activity of this gene has been associated with human cancer. Alternatively spliced transcript variants have been observed. The use of alternative translation start sites results in isoforms with longer or shorter N-termini which may differ in their subcellular localizations. There are two pseudogenes for this gene on chromosome 17. [provided by RefSeq, Jan 2013].

Function : Serine/threonine-protein kinase that acts downstream of mTOR signaling in response to growth factors and nutrients to promote cell proliferation, cell growth and cell cycle progression. Regulates protein synthesis through phosphorylation of EIF4B, RPS6 and EEF2K, and contributes to cell survival by repressing the pro-apoptotic function of BAD. Under conditions of nutrient depletion, the inactive form associates with the EIF3 translation initiation complex. Upon mitogenic stimulation, phosphorylation by the mammalian target of rapamycin complex 1 (mTORC1) leads to dissociation from the EIF3 complex and activation. The active form then phosphorylates and activates several substrates in the preinitiation complex, including the EIF2B complex and the cap-binding complex component EIF4B. Also controls translation initiation by phosphorylating a negative regulator of EIF4A, PDCD4, targeting it for ubiquitination and subsequent proteolysis. Promotes initiation of the pioneer round of protein synthesis by phosphorylating POLDIP3/SKAR. In response to IGF1, activates translation elongation by phosphorylating EEF2 kinase (EEF2K), which leads to its inhibition and thus activation of EEF2. Also plays a role in feedback regulation of mTORC2 by mTORC1 by phosphorylating RICTOR, resulting in the inhibition of mTORC2 and AKT1 signaling. Mediates cell survival by phosphorylating the pro-apoptotic protein BAD and suppressing its pro-apoptotic function. Phosphorylates mitochondrial URI1 leading to dissociation of a URI1-PPP1CC complex. The free mitochondrial PPP1CC can then dephosphorylate RPS6KB1 at 'Thr-412', which is proposed to be a negative feedback mechanism for the RPS6KB1 anti-apoptotic function. Mediates TNF-alpha-induced insulin resistance by phosphorylating IRS1 at multiple serine residues, resulting in accelerated degradation of IRS1. In cells lacking functional TSC1-2 complex, constitutively phosphorylates and inhibits GSK3B. May be involved in cytoskeletal rearrangement through binding to neurabin.

Subunit : Interacts with PPP1R9A/neurabin-1. Interacts with RPTOR. Interacts with IRS1. Interacts with EIF3B and EIF3C. Interacts with POLDIP3 and TRAF4.

Subcellular Location : Cell junction, synapse, synaptosome. Mitochondrion outer membrane. Mitochondrion.Note=Colocalizes with URI1 at mitochondrion. Isoform Alpha I: Nucleus. Cytoplasm. Isoform Alpha II: Cytoplasm.

Post-translational modifications : Phosphorylation at Thr-412 is regulated by mTORC1. The phosphorylation at this site is maintained by an agonist-dependent autophosphorylation mechanism (By similarity). Activated by phosphorylation at Thr-252 by PDPK1. Dephosphorylation by PPP1CC at Thr-412 in mitochondrion.

Similarity : Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. S6 kinase subfamily.

Contains 1 AGC-kinase C-terminal domain.

Contains 1 protein kinase domain.

Database links : UniProtKB/Swiss-Prot: P23443.2

抗体的鉴定：

1）磷酸化核糖体S6蛋白激酶β2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-ICAM-2/CD102 /FITC 荧光素标记细胞间粘附分子-2抗体IgGMulti-class antibodies规格： 0.2ml

Anti-IGSF8/CD3/FITC 荧光素标记免疫球蛋超家族成员8抗体IgGMulti-class antibodies规格： 0.2ml

Rhesus antibody Rh CD44/HCAM/PGP1 CD44抗体 规格 0.1ml

Notch1/MOTC(Neurogenic locus notch homolog protein 1) 跨膜受体蛋白Notch-1抗原 0.5mg

IHH 英文名称： HHG2抗体 0.2ml

Rhesus antibody Rh Synaptotagmin 5 突触结合蛋白5抗体 规格 0.2ml

Anti-IGSF8/CD3/FITC 荧光素标记免疫球蛋超家族成员8抗体IgGMulti-class antibodies规格： 0.2ml

Alpha-SYN(Human Alpha-Synuclein) ELISA Kit 人α突触核蛋白Multi-class antibodies规格： 48T

Anti-Phospho-PLC beta3 (Ser1105) 磷酸化磷酯酶Cβ3抗体Multi-class antibodies规格： 0.1ml

Rhesus antibody Rh HIFPH4 缺氧诱导因子脯氨酰4羟化酶抗体 规格 0.2ml

CsA(Mouse Cyclosporine A) ELISA Kit 小鼠环孢素A 96T

Properdin 英文名称： P因子/备解素抗体 0.1ml

CD161 英文名称： CD161抗体 0.1ml

Anti-Phospho-PLC beta3 (Ser1105) 磷酸化磷酯酶Cβ3抗体Multi-class antibodies规格： 0.1ml

PAF ELISA Kit 大鼠血小板活化因子Multi-class antibodies规格： 48T

Anti-Adipo R2 /FITC 荧光素标记脂联素受体-2抗体IgGMulti-class antibodies规格： 0.2ml

Rhesus antibody Rh Fetuin beta 胎球蛋白B抗体(Fetuin β) 规格 0.2ml

Lep IgG(Mouse Leptospira IgG) ELISA Kit 小鼠钩端螺旋体IgG 96T

Phospho-MEK1/2 (Ser218 + Ser222) 英文名称： 磷酸化原活化蛋白激酶激酶1/2抗体 0.1ml

ARRDC1 英文名称： 抑制蛋白结构域蛋白1抗体 0.2ml

Anti-Adipo R2 /FITC 荧光素标记脂联素受体-2抗体IgGMulti-class antibodies规格： 0.2ml

293来源病毒包装细胞;ΦA-GP

FST Others Human 人 FST / Follistatin 人细胞裂解液 (阳性对照)

NCI-H524 人小细胞

mCF, 小鼠成纤维细胞

人骨髓间充质干细胞(hMSCs)

人;HFL1 人淋巴成纤维细胞完全培养基 100mL

293来源病毒包装细胞;ΦA-GP

FST Others Human 人 FST / Follistatin 人细胞裂解液 (阳性对照)

NCI-H524 人小细胞

mCF, 小鼠成纤维细胞

人骨髓间充质干细胞(hMSCs)

人;HFL1 人淋巴成纤维细胞完全培养基 100mL

磷酸化核糖体S6蛋白激酶β2抗体说明书 LTA4H Others Human 人 Leukoiene A4 Hydrolase / LTA4H 杆状病毒-昆虫细胞裂解液 (阳性对照)

CL-0056CCD-1095Sk(人浸润性导管癌旁皮肤细胞)5×106cells/瓶×2

β-半乳糖苷酶比色检测试剂盒GalC

Cates-1B细胞，人淋巴胚胎性癌细胞 SV40T转化的人胚肾细胞(亚系),293T/17细胞 食管上皮细胞Many types of cells包装：5 × 105次方(1ml)

大耳山羊肺细胞;LDG-1

SFRP1 Others Mouse 小鼠 sFRP1 人细胞裂解液 (阳性对照)