抗體的生物素化標(biāo)記實驗要點：

1. 磷酸化核糖體S6蛋白激酶β2抗體說明書 如在反應(yīng)混合液中有疊氮鈉或游離氨基存在,會抑制標(biāo)記反應(yīng)。因此,蛋白質(zhì)在反應(yīng)前要對 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-氨基的密度會有所不同,選擇不當(dāng)則影響標(biāo)記的效率,應(yīng)先用幾個不同的分子比來篩選最適條件；

3.用NHSB量過量也是不利的,抗原的結(jié)合位點可能因此被封閉,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,此時可加入去污劑如 Triton x-100, Tween20等；

5.當(dāng)游離ε-氨基(賴氨酸殘基的氨基)存在于抗體的抗原結(jié)合位點時,或位于酶的催化位點時,生物素化會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,如羰基、氨基、巰基、異咪唑基及苯酚基,也可與糖基共價結(jié)合；

7.交聯(lián)反應(yīng)后,應(yīng)充分透析,否則,殘余的生物素會對生物素化抗體與親和素的結(jié)合產(chǎn)生競爭作用；

8.在細胞的熒光標(biāo)記實驗中,中和親和素的本底低,但由于鏈霉親和素含有少量正電荷,故對某些細胞可導(dǎo)致高本底。

產(chǎn)品訂購信息：
英文名稱  Anti-Phospho-p70 S6 Kinase Beta (Thr444 + Ser447) 

中文名稱   磷酸化核糖體S6蛋白激酶β2抗體說明書 

別    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.

濃    度   1mg/1ml

規(guī) 格  0.1ml/100μg

抗體來源   Rabbit

克隆類型   polyclonal

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

產(chǎn)品類型   一抗 磷酸化抗體  

研究領(lǐng)域     免疫學(xué) 信號轉(zhuǎn)導(dǎo) 細胞凋亡 轉(zhuǎn)錄調(diào)節(jié)因子 激酶和磷酸酶

蛋白分子量  predicted molecular weight: 70kDa

性    狀   Lyophilized or Liquid

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

亞    型   IgG

純化方法   affinity purified by Protein A

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

磷酸化核糖體S6蛋白激酶β2抗體說明書 產(chǎn)品應(yīng)用    WB=1:100-500 ELISA=1:500-1000 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 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

抗體的鑒定：

1）磷酸化核糖體S6蛋白激酶β2抗體說明書 抗體的效價鑒定：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。不同的抗原制備的抗體,要求的效價不一。鑒定效價的方法很多,包括有試管凝集反應(yīng),瓊脂擴散試驗,酶聯(lián)免疫吸附試驗等。常用的抗原所制備的抗體一般都有約成的鑒定效價的方法,以資比較。如制備抗抗體的效價,一般就采用瓊脂擴散試驗來鑒定。

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)的識別能力?？贵w的特異性高,它的識別能力就強。衡量特異性通常以交叉反應(yīng)率來表示。交叉反應(yīng)率可用競爭抑制試驗測定。以不同濃度抗原和近似抗原分別做競爭抑制曲線,計算各自的結(jié)合率,求出各自在IC50時的濃度,并按公式計算交叉反應(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濃度幾乎是無窮大時,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親和力的測定對抗體的篩選,確定抗體的用途,驗證抗體的均一性等均有重要意義。

Anti-ICAM-2/CD102 /FITC 熒光素標(biāo)記細胞間粘附分子-2抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Anti-IGSF8/CD3/FITC 熒光素標(biāo)記免疫球蛋超家族成員8抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh CD44/HCAM/PGP1 CD44抗體 規(guī)格 0.1ml

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

IHH 英文名稱： HHG2抗體 0.2ml

Rhesus antibody Rh Synaptotagmin 5 突觸結(jié)合蛋白5抗體 規(guī)格 0.2ml

Anti-IGSF8/CD3/FITC 熒光素標(biāo)記免疫球蛋超家族成員8抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Alpha-SYN(Human Alpha-Synuclein) ELISA Kit 人α突觸核蛋白Multi-class antibodies規(guī)格： 48T

Anti-Phospho-PLC beta3 (Ser1105) 磷酸化磷酯酶Cβ3抗體Multi-class antibodies規(guī)格： 0.1ml

Rhesus antibody Rh HIFPH4 缺氧誘導(dǎo)因子脯氨酰4羥化酶抗體 規(guī)格 0.2ml

CsA(Mouse Cyclosporine A) ELISA Kit 小鼠環(huán)孢素A 96T

Properdin 英文名稱： P因子/備解素抗體 0.1ml

CD161 英文名稱： CD161抗體 0.1ml

Anti-Phospho-PLC beta3 (Ser1105) 磷酸化磷酯酶Cβ3抗體Multi-class antibodies規(guī)格： 0.1ml

PAF ELISA Kit 大鼠血小板活化因子Multi-class antibodies規(guī)格： 48T

Anti-Adipo R2 /FITC 熒光素標(biāo)記脂聯(lián)素受體-2抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh Fetuin beta 胎球蛋白B抗體(Fetuin β) 規(guī)格 0.2ml

Lep IgG(Mouse Leptospira IgG) ELISA Kit 小鼠鉤端螺旋體IgG 96T

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

ARRDC1 英文名稱： 抑制蛋白結(jié)構(gòu)域蛋白1抗體 0.2ml

Anti-Adipo R2 /FITC 熒光素標(biāo)記脂聯(lián)素受體-2抗體IgGMulti-class antibodies規(guī)格： 0.2ml

293來源病毒包裝細胞;ΦA(chǔ)-GP

FST Others Human 人 FST / Follistatin 人細胞裂解液 (陽性對照)

NCI-H524 人小細胞

mCF, 小鼠成纖維細胞

人骨髓間充質(zhì)干細胞(hMSCs)

人;HFL1 人淋巴成纖維細胞完全培養(yǎng)基 100mL

293來源病毒包裝細胞;ΦA(chǔ)-GP

FST Others Human 人 FST / Follistatin 人細胞裂解液 (陽性對照)

NCI-H524 人小細胞

mCF, 小鼠成纖維細胞

人骨髓間充質(zhì)干細胞(hMSCs)

人;HFL1 人淋巴成纖維細胞完全培養(yǎng)基 100mL

磷酸化核糖體S6蛋白激酶β2抗體說明書 LTA4H Others Human 人 Leukoiene A4 Hydrolase / LTA4H 桿狀病毒-昆蟲細胞裂解液 (陽性對照)

CL-0056CCD-1095Sk(人浸潤性導(dǎo)管癌旁皮膚細胞)5×106cells/瓶×2

β-半乳糖苷酶比色檢測試劑盒GalC

Cates-1B細胞，人淋巴胚胎性癌細胞 SV40T轉(zhuǎn)化的人胚腎細胞(亞系),293T/17細胞 食管上皮細胞Many types of cells包裝：5 × 105次方(1ml)

大耳山羊肺細胞;LDG-1

SFRP1 Others Mouse 小鼠 sFRP1 人細胞裂解液 (陽性對照)