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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) | CS12157 |
| 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-STAT5b(Ser731) |
| 克隆性 | 是 |
| 靶點(diǎn) | 詳見(jiàn)說(shuō)明書(shū) |
| 適應(yīng)物種 | 詳見(jiàn)說(shuō)明書(shū) |
| 形態(tài) | 詳見(jiàn)說(shuō)明書(shū) |
| 宿主 | 詳見(jiàn)說(shuō)明書(shū) |
| 亞型 | IgG |
| 標(biāo)識(shí)物 | 詳見(jiàn)說(shuō)明書(shū) |
| 濃度 | 1mg/1ml% |
| 免疫原 | KLH conjugated Synthesised phosphopeptide derived from human STAT5b around the phosphorylation site of Ser731 |
抗體的生物素化標(biāo)記實(shí)驗(yàn)要點(diǎn)：
1. 磷酸化信號(hào)轉(zhuǎn)導(dǎo)和轉(zhuǎn)錄激活因子5b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。
產(chǎn)品訂購(gòu)信息：
英文名稱 Anti-phospho-STAT5b(Ser731)
中文名稱 磷酸化信號(hào)轉(zhuǎn)導(dǎo)和轉(zhuǎn)錄激活因子5b抗體費(fèi)用
別 名 STAT5b(phospho S731); Transcription factor STAT5B; Signal transducer and activator of transcription 5B; STA5B_HUMAN; STAT5; Stat5b.
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, Chicken, Dog, Pig, Cow, Horse, Sheep
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) 染色質(zhì)和核信號(hào) 信號(hào)轉(zhuǎn)導(dǎo) 轉(zhuǎn)錄調(diào)節(jié)因子
蛋白分子量 predicted molecular weight: 90kDa
性 狀 Lyophilized or Liquid
免 疫 原 KLH conjugated Synthesised phosphopeptide derived from human STAT5b around the phosphorylation site of Ser731
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
磷酸化信號(hào)轉(zhuǎn)導(dǎo)和轉(zhuǎn)錄激活因子5b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 Carries out a dual function: signal transduction and activation of transcription. Binds to the GAS element and activates PRL-induced transcription.
Function : Carries out a dual function: signal transduction and activation of transcription. Mediates cellular responses to the cytokine KITLG/SCF and other growth factors. Binds to the GAS element and activates PRL-induced transcription.
Subunit : Forms a homodimer or a heterodimer with a related family member. Binds NR3C1 (By similarity). Interacts with NCOA1, NMI and SOCS7. Interacts (via SH2 domain) with INSR.
Subcellular Location : Cytoplasm (By similarity). Nucleus (By similarity). Note=Translocated into the nucleus in response to phosphorylation (By similarity).
Post-translational modifications : Tyrosine phosphorylated in response to signaling via activated KIT, resulting in translocation to the nucleus. Tyrosine phosphorylated in response to signaling via activated FLT3; wild-type FLT3 results in much weaker phosphorylation than constitutively activated mutant FLT3. Alternatively, can be phosphorylated by JAK2. Phosphoryation at Tyr-699 by PTK6 or HCK leads to an increase of its transcriptional activity.
DISEASE : Growth hormone insensitivity with immunodeficiency (GHII) [MIM:245590]: A disease characterized by short stature, growth hormone deficiency in the presence of normal to elevated circulating concentrations of growth hormone, resistance to hexogeneous growth hormone therapy, and recurrent infections. Note=The disease is caused by mutations affecting the gene represented in this entry.
Similarity : Belongs to the transcription factor STAT family.
Contains 1 SH2 domain.
Database links : UniProtKB/Swiss-Prot: P51692.2
抗體的鑒定：
1）磷酸化信號(hào)轉(zhuǎn)導(dǎo)和轉(zhuǎn)錄激活因子5b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。
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