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產(chǎn)地 | 進(jìn)口、國(guó)產(chǎn) |
品牌 | 上海莼試 |
保存條件 | Store at -20 °C |
貨號(hào) | CS10245 |
應(yīng)用范圍 | 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 |
CAS編號(hào) | |
抗體名 | Anti-phospho-PKA R2 (Ser96) |
克隆性 | |
靶點(diǎn) | 詳見(jiàn)說(shuō)明書 |
適應(yīng)物種 | 詳見(jiàn)說(shuō)明書 |
形態(tài) | 詳見(jiàn)說(shuō)明書 |
宿主 | 詳見(jiàn)說(shuō)明書 |
亞型 | IgG |
標(biāo)識(shí)物 | 詳見(jiàn)說(shuō)明書 |
濃度 | 1mg/1ml% |
免疫原 | KLH conjugated Synthesised phosphopeptide derived from mouse PKA R2 around the phosphorylation site of Ser96 |
產(chǎn)品訂購(gòu)信息:
英文名稱 Anti-phospho-PKA R2 (Ser96)
中文名稱 磷酸化蛋白激酶A受體2α亞基抗體費(fèi)用
別 名 KAP2; KAP2_HUMAN; MGC3606; PKR 2; PKR2; PRKA R2; PRKAR 2; PRKAR2; PRKAR2A; Protein kinase A RII alpha subunit; Protein kinase cAMP dependent regulatory type II alpha; cAMP dependent protein kinase regulatory subunit alpha 2; cAMP dependent protein kinase regulatory subunit RII alpha; cAMP dependent protein kinase type II alpha regulatory chain; cAMP dependent protein kinase type II alpha regulatory subunit; cAMP-dependent protein kinase type II-alpha regulatory subunit; phospho-PKA R2 (Ser98)(human).
濃 度 1mg/1ml
規(guī) 格 0.1ml/100μg
抗體來(lái)源 Rabbit
克隆類型 polyclonal
交叉反應(yīng) Human, Mouse, Rat
產(chǎn)品類型 一抗 磷酸化抗體
研究領(lǐng)域 細(xì)胞生物 免疫學(xué) 轉(zhuǎn)錄調(diào)節(jié)因子
蛋白分子量 predicted molecular weight: 45kDa
性 狀 Lyophilized or Liquid
免 疫 原 KLH conjugated Synthesised phosphopeptide derived from mouse PKA R2 around the phosphorylation site of Ser96
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A受體2α亞基抗體費(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 The second messenger cyclic AMP (cAMP) mediates diverse cellular responses to external signals such as proliferation, ion transport, regulation of metabolism and gene transcription by activation of the cAMP-dependent protein kinase (cAPK or PKA). Activation of PKA occurs when cAMP binds to the two regulatory subunits of the tetrameric PKA holoenzyme, resulting in release of active catalytic subunits. Activation of transcription upon elevation of cAMP levels results from translocation of PKA to the nucleus, where it phosphorylates the transcription factor cAMP response element binding protein (CREB) on Serine 133, which in turn leads to TFIIB binding to TATA-box-binding protein TBP1, thus linking phospho-CREB to the Pol II transcription initiation complex. Mouse Serine 96 (designated Ser 99 in human) is a phosphorylation site on the PKA II?regulatory subunit.
Function : Regulatory subunit of the cAMP-dependent protein kinases involved in cAMP signaling in cells. Type II regulatory chains mediate membrane association by binding to anchoring proteins, including the MAP2 kinase.
Subunit : The inactive form of the enzyme is composed of two regulatory chains and two catalytic chains. Activation by cAMP produces two active catalytic monomers and a regulatory dimer that binds four cAMP molecules. Interacts with AKAP4 and CBFA2T3. Interacts with the phosphorylated form of PJA2.
Subcellular Location : Cytoplasm. Cell membrane. Note=Co-localizes with PJA2 in the cytoplasm and the cell membrane.
Post-translational modifications : Phosphorylated by the activated catalytic chain.
Similarity : Belongs to the cAMP-dependent kinase regulatory chain family.
Database links : UniProtKB/Swiss-Prot: P13861.2
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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A受體2α亞基抗體費(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(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*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。
抗體的鑒定:
1)磷酸化蛋白激酶A受體2α亞基抗體費(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(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,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。