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產(chǎn)地 | 進(jìn)口、國產(chǎn) |
品牌 | 上海莼試 |
保存條件 | Store at -20 °C |
貨號(hào) | CS10592 |
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-PAR4 |
克隆性 | |
靶點(diǎn) | 詳見說明書 |
適應(yīng)物種 | 詳見說明書 |
形態(tài) | 詳見說明書 |
宿主 | 詳見說明書 |
亞型 | IgG |
標(biāo)識(shí)物 | 詳見說明書 |
濃度 | 1mg/1ml% |
免疫原 | KLH conjugated synthetic peptide derived from human PAR4 C-terminus |
抗體的生物素化標(biāo)記實(shí)驗(yàn)要點(diǎn):
1. 蛋白酶活化受體4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(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。
產(chǎn)品訂購信息:
英文名稱 Anti-PAR4
中文名稱 蛋白酶活化受體4抗體說明書
別 名 protease-activated receptor 4; 2310001G03Rik; PAR 4; Par4 induced by effectors of apoptosis; PAWR; PRKC Apoptosis WT1 Regulator; PRKC apoptosis WT1 regulator protein; Prostate apoptosis response 4 protein; Prostate apoptosis response protein 4; Prostate apoptosis response protein PAR 4; Proteinase-Activated Receptor 4; Transcriptional repressor Par-4-like protein PAWR; TRANSCRIPTIONAL REPRESSOR PAR4; WT1 Interacting Protein; PAR-4.
濃 度 1mg/1ml
規(guī) 格 0.1ml/100μg 0.2ml/200μg
抗體來源 Rabbit
克隆類型 polyclonal
交叉反應(yīng) Human, Mouse, Rat, Dog, Pig, Cow, Horse, Guinea Pig
產(chǎn)品類型 一抗
研究領(lǐng)域 細(xì)胞生物 信號(hào)轉(zhuǎn)導(dǎo) 激酶和磷酸酶 細(xì)胞膜受體
蛋白分子量 predicted molecular weight: 37kDa
性 狀 Lyophilized or Liquid
免 疫 原 KLH conjugated synthetic peptide derived from human PAR4 C-terminus
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
蛋白酶活化受體4抗體說明書 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 PAR4 is a member of a unique family of GPCRs, the proteinase-activated receptors (PARs), that are activated by proteolytic cleavage of the N-terminal domain of the receptor to reveal a tethered ligand. The PAR family consists of 4 receptors; PAR1 and PAR3 are activated by thrombin, and PAR2 and PAR4 are activated by several serine proteases (Macfarlane et al., 2001). PAR4 is expressed along with PAR1 on human platelets, and each plays an important role in thrombin-induced platelet aggregation (Holinstat et al., 2006). However, platelets from mice express PAR4 but not PAR1, and platelets from mice lacking PAR4 fail to aggregate in response to thrombin (Sambrano et al., 2001). Mice lacking PAR4 are protected from mesenteric arteriole thrombosis, indicating that PAR4 is a potential target for treatment of thrombosis in humans.
Function : Receptor for activated thrombin or trypsin coupled to G proteins that stimulate phosphoinositide hydrolysis. May play a role in platelets activation.
Subcellular Location : Cell membrane; Multi-pass membrane protein.
Tissue Specificity : Widely expressed, with highest levels in lung, pancreas, thyroid, testis and small intestine. Not expressed in brain, kidney, spinal cord and peripheral blood leukocytes. Also detected in platelets.
Similarity : Belongs to the G-protein coupled receptor 1 family.
蛋白酶活化受體-4(細(xì)胞凋亡反應(yīng)蛋白4)是蛋白酶活化受體PARs (protease-activated receptors , PARs)的成員之一,PAR4在激活、滅活、脫敏、復(fù)敏、及其與信號(hào)轉(zhuǎn)導(dǎo)途徑的關(guān)系,尤其是與疾病的關(guān)系正倍受關(guān)注。
PAR4具有廣泛的生物學(xué)效應(yīng),最終影響細(xì)胞的形態(tài)的改變、分泌蛋白和整合蛋白的活化、代謝反應(yīng)、轉(zhuǎn)錄過程和細(xì)胞運(yùn)動(dòng),由此參與細(xì)胞的信號(hào)轉(zhuǎn)導(dǎo)作用。
抗體的鑒定:
1)蛋白酶活化受體4抗體說明書 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。
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