產(chǎn)品訂購信息：
英文名稱  Anti-Phospho-PKR (Thr446/451) 

中文名稱   磷酸化蛋白激酶R抗體費(fèi)用 

別    名  double-stranded RNA-dependent Protein Kinase; interferon-induced, double-stranded RNA-activated protein kinase isoform a; protein kinase, interferon-inducible double stranded RNA dependent; interferon-inducible elF2alpha kinase; double stranded RNA activated protein kinase; p68 kinase; eIF-2A protein kinase 2; P1/eIF-2A protein kinase; protein kinase RNA-activated; interferon-inducible RNA-dependent protein kinase; EIF2AK2; EIF2AK1; MGC126524; PKR; PRKR; E2AK2_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 

產(chǎn)品類型   一抗 磷酸化抗體  

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

蛋白分子量  predicted molecular weight: 62kDa

性    狀   Lyophilized or Liquid

免 疫 原 KLH conjugated synthesised phosphopeptide derived from human PKR around the phosphorylation site of Thr446/451 

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

磷酸化蛋白激酶R抗體費(fèi)用 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 PKR is an interferon-inducible serine/threonine specific protein kinase. It is widely expressed in eukaryotic organisms and activated by double stranded RNA. Activation of PKR by dsRNAs leads to autophosphorylation at multiple sites. Phosphorylation of Thr446 and Thr451 in the PKR activation loop is required in vivo and in vitro for high level kinase activity. PKR phosphorylates its natural substrate, the alpha subunit of eukaryotic protein synthesis initiation factor 2 (EIF2 alpha), leading to the inhibition of protein synthesis. PKR is also involved in TLR signaling and mediates apoptosis in fibroblasts in response to viral infection and inflammatory cytokines, and also activates IKK and NFKB, thereby suppressing apoptosis. Recently, it has been reported that PKR also phosphorylates human p53 on serine 392. PKR might play a role in ER stress-induced apoptosis and in Alzheimer's disease. Alzheimer cases show prominent PKR activation in association with neuritic plaques and pyramidal neurons in the hippocampus and neocortex.

Function : Following activation by double-stranded RNA in the presence of ATP, the kinase becomes autophosphorylated and can catalyze the phosphorylation of the translation initiation factor EIF2S1, which leads to an inhibition of the initiation of protein synthesis. Double-stranded RNA is generated during the course of a viral infection. In adddition to serine/threonine-protein kinase activity, also has tyrosine-protein kinase activity: phosphorylates CDK1 upon DNA damage. CDK1 phosphorylation triggers CDK1 polyubiquitination and subsequent proteolysis, thus leading to G2 arrest

Subunit : Homodimer. Interacts with STRBP (By similarity). Interacts with DNAJC3. Inhibited by direct interaction with viral proteins such as HCV E2, HCV NS* and influenza A NS1. Activated by the interaction with HIV-1 Tat (By similarity). Forms a complex with FANCA, FANCC, FANCG and HSP70.

Post-translational modifications : Autophosphorylated on several Ser and Thr residues. Autophosphorylation of Thr-451 is dependent on Thr-446 and is stimulated by dsRNA binding and dimerization. Autophosphorylation apparently leads to the activation of the kinase.

Contains 2 DRBM (double-stranded RNA-binding) domains.

Contains 1 protein kinase domain.

Database links : UniProtKB/Swiss-Prot: P19525.2

Anti-CD133/Biotin 生物素化造血干細(xì)胞抗原CD133抗體Multi-class antibodies規(guī)格： 0.2ml

Anti-ID1/Inhibitor of DNA binding 1 /FITC 熒光素標(biāo)記DNA結(jié)合抑制因子1抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh CD57/HNK1 髓鞘相關(guān)糖蛋白CD57抗體 規(guī)格 0.1ml

MKK3(MAP Kinase Kinase 3) 原活化蛋白激酶MKK3/6抗原 0.5mg

IFI-202 英文名稱： γ干擾素誘導(dǎo)蛋白202抗體 0.2ml

Rhesus antibody Rh TACC1 癌、抗原GA55抗體 規(guī)格 0.2ml

Anti-ID1/Inhibitor of DNA binding 1 /FITC 熒光素標(biāo)記DNA結(jié)合抑制因子1抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Anti-ISR/FITC 熒光素標(biāo)記受體抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Anti-Phospho-SGK1 (Ser422) /FITC 熒光素標(biāo)記磷酸化糖皮質(zhì)激素調(diào)節(jié)激酶1抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh Androgen receptor 雄激素受體抗體 規(guī)格 0.1ml

Rabbit Anti-Monkey IgG/FITC FITC標(biāo)記的兔抗猴IgG 0.3ml

GP210 英文名稱： 核膜糖蛋白210抗體 0.2ml

Rhesus antibody Rh Rabbit Anti-horse IgM/Alexa Fluor 488 Alexa Fluor 488標(biāo)記的兔抗馬IgM 規(guī)格 0.1ml

Anti-Phospho-SGK1 (Ser422) /FITC 熒光素標(biāo)記磷酸化糖皮質(zhì)激素調(diào)節(jié)激酶1抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Amylin ELISA Kit 大鼠胰淀素Multi-class antibodies規(guī)格： 48T

Anti-phospho-VEGF receptor 2 (Tyr951) 磷酸化內(nèi)皮生長因子受體2抗體Multi-class antibodies規(guī)格： 0.1ml

Rhesus antibody Rh GALT 磷酸半乳苷酸轉(zhuǎn)移酶1抗體 規(guī)格 0.2ml

ECF(Human eosinophil chemotactic factor) ELISA Kit 人鼠嗜酸粒細(xì)胞趨化因子 96T

Phospho-NMDAR2B (Tyr1336) 英文名稱： 磷酸化谷酸受體2B抗體 0.1ml

phospho-ADRB2 (Ser261) 英文名稱： 磷酸化能受體β2抗體 0.1ml

Anti-phospho-VEGF receptor 2 (Tyr951) 磷酸化內(nèi)皮生長因子受體2抗體Multi-class antibodies規(guī)格： 0.1ml

CL-0114Hs 578T(人癌細(xì)胞)5×106cells/瓶×2

KIR2DL4 Others Human 人 KIR2DL4 / CD158D CHO細(xì)胞裂解液 (陽性對(duì)照)

人外根殼細(xì)胞裂解物HHORSCL

牛源細(xì)胞 細(xì)胞，HCCLM3細(xì)胞 C3細(xì)胞，小鼠細(xì)胞

人胚腎細(xì)胞(SV40T基因修飾);293T/17

COCH Others Human 人 COCH / Cochlin 人細(xì)胞裂解液 (陽性對(duì)照)

CL-0112HMy2.CIR(人B淋巴母細(xì)胞)5×106cells/瓶×2

DDC Others Human 人 DOPA Decarboxylase / DDC 桿狀病毒-昆蟲細(xì)胞裂解液 (陽性對(duì)照)

人毛細(xì)胞裂解物HHDPCL

EB-3[EB3]細(xì)胞，人淋巴樣Butt細(xì)胞 人腎透明細(xì)胞腺癌細(xì)胞,786-O細(xì)胞 前脂肪細(xì)胞Many types of cells包裝：5 × 105次方(1ml)

兔間變表皮鱗株;VX2

SPI2 Others Mouse 小鼠 SPI2 / HAI2 人細(xì)胞裂解液 (陽性對(duì)照)

磷酸化蛋白激酶R抗體費(fèi)用 HA Others H1N1 甲型 H1N1 (A/New York/18/2009) HA 人細(xì)胞裂解液 (陽性對(duì)照)

人*上皮細(xì)胞HREpiC

IEC-6 大鼠小腸引窩上皮細(xì)胞

CM-H115人腦動(dòng)脈內(nèi)皮細(xì)胞完全培養(yǎng)基100mL

U266B1 [U266], 人細(xì)胞

人細(xì)胞;Hela 人晶狀體上皮細(xì)胞完全培養(yǎng)基 100mL

抗體的生物素化標(biāo)記實(shí)驗(yàn)要點(diǎn)：

1. 磷酸化蛋白激酶R抗體費(fèi)用 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(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)生競爭作用；

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。

抗體的鑒定：

1）磷酸化蛋白激酶R抗體費(fèi)用 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。抗體的特異性高,它的識(shí)別能力就強(qiáng)。衡量特異性通常以交叉反應(yīng)率來表示。交叉反應(yīng)率可用競爭抑制試驗(yàn)測定。以不同濃度抗原和近似抗原分別做競爭抑制曲線,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(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。