抗體的生物素化標記實驗要點：

1. 凝血酶(凝血因子II)激活肽1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,選擇不當則影響標記的效率,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-Thrombin Activation peptide fragment 1 

中文名稱   凝血酶(凝血因子II)激活肽1抗體說明書 

別    名  Activation peptide fragment 1; coagulation factor II; prothrombin; F2; Cf-2; Cf2; FII; F 2; coagulation factor II (thrombin); Coagulation factor II; Coagulation factor II precursor; F2; Factor II; Factor-II; Prothrombin; prothrombin B-chain; PT; serine protease; THRB; THRB_HUMAN; Thrombin.

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, Guinea Pig 

產(chǎn)品類型   一抗  

研究領(lǐng)域    心 細胞生物 激酶和磷酸酶 合成與降解

蛋白分子量  predicted molecular weight: 17/64kDa

性    狀   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human Thrombin Activation peptide fragment 1 

亞    型   IgG

純化方法   affinity purified by Protein A

儲 存 液   Preservative: 15mM Sodium Azide, Constituents: 1% BSA, 0.01M PBS, pH 7.4

凝血酶(凝血因子II)激活肽1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 Thrombin is the final protease in the blood coagulation cascade and serves both pro- and anticoagulant functions through the cleavage of several targets. The ability of thrombin to specifically recognize a wide range of substrates derives from interactions which occur outside of the active site of thrombin. Thrombin possesses two anion binding exosites which mediate many of its interactions with cofactors and substrates, and although many structures of thrombin have been solved, few such interactions have been described in molecular detail. Glycosaminoglycan binding to exosite II of thrombin plays a major role in switching off the procoagulant functions of thrombin by mediating its irreversible inhibition by circulating serpins and by its binding to the endothelial cell surface receptor thrombomodulin.

Function : Thrombin is an active enzyme in the earliest steps of the blood clot formation, generated from its circulating inactive precursor prothrombin. Thrombin is a glycoprotein formed by two peptides chains of 36 and 259 amino acids linked by disulfure bonds. Three important sites have been identified on the surface of the enzyme: The catalytic site that confers to the molecule its serine protease activity, the exosite one responsible for the binding of the substrate (fibrinogen or thrombin receptor) and the exosite two responsible for the binding of antithrombin III and inactivation of thrombin. Gamma Thrombin is a proteolyzed form of Alpha Thrombin. Gamma Thrombin consists of four chains (A, B1, B5, B4) with a disulfide link between the A peptide and the B5 peptide.

Subunit : Heterodimer (named alpha-thrombin) of a light and a heavy chain; disulfide-linked. Forms a heterodimer with SERPINA5.

Subcellular Location : Secreted, extracellular space.

Tissue Specificity : Expressed by the liver and secreted in plasma.

Post-translational modifications : The gamma-carboxyglutamyl residues, which bind calcium ions, result from the carboxylation of glutamyl residues by a microsomal enzyme, the vitamin K-dependent carboxylase. The modified residues are necessary for the calcium-dependent interaction with a negatively charged phospholipid surface, which is essential for the conversion of prothrombin to thrombin.

N-glycosylated. N-glycan heterogeneity at Asn-121: Hex3HexNAc3 (minor), Hex4HexNAc3 (minor) and Hex5HexNAc4 (major). At Asn-143: Hex4HexNAc3 (minor) and Hex5HexNAc4 (major).

DISEASE : Factor II deficiency (FA2D) [MIM:613679]: A very rare blood coagulation disorder characterized by mucocutaneous bleeding symptoms. The severity of the bleeding manifestations correlates with blood factor II levels. Note=The disease is caused by mutations affecting the gene represented in this entry.

Ischemic stroke (ISCHSTR) [MIM:601367]: A stroke is an acute neurologic event leading to death of neural tissue of the brain and resulting in loss of motor, sensory and/or cognitive function. Ischemic strokes, resulting from vascular occlusion, is considered to be a highly complex disease consisting of a group of heterogeneous disorders with multiple genetic and environmental risk factors. Note=Disease susceptibility is associated with variations affecting the gene represented in this entry.

Thrombophilia due to thrombin defect (THPH1) [MIM:188050]: A multifactorial disorder of hemostasis characterized by abnormal platelet aggregation in response to various agents and recurrent thrombi formation. Note=The disease is caused by mutations affecting the gene represented in this entry. A common genetic variation in the 3-prime untranslated region of the prothrombin gene is associated with elevated plasma prothrombin levels and an increased risk of venous thrombosis.

Pregnancy loss, recurrent, 2 (RPRGL2) [MIM:614390]: A common complication of pregnancy, resulting in spontaneous abortion before the fetus has reached viability. The term includes all miscarriages from the time of conception until 24 weeks of gestation. Recurrent pregnancy loss is defined as 3 or more consecutive spontaneous abortions. Note=Disease susceptibility is associated with variations affecting the gene represented in this entry.

Similarity : Belongs to the peptidase S1 family.

Contains 1 Gla (gamma-carboxy-glutamate) domain.

Contains 2 kringle domains.

Contains 1 peptidase S1 domain.

Database links : UniProtKB/Swiss-Prot: P00734.2

抗體的鑒定：

1）凝血酶(凝血因子II)激活肽1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。抗體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-SLK/Fyn/FITC 熒光素標記酪氨酸蛋白激酶Fyn(同步原癌基因)抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Caspase-1 (ICE; CASP-1;P45) 天冬氨酸-胱氨酸特異性蛋白酶家族(抗原)Multi-class antibodies規(guī)格： 0.5mg

核因子2相關(guān)因子2抗體 Anti-Nrf2 0.1ml

pan 14-3-3 英文名稱： 14-3-3蛋白抗體 0.2ml

EMP3 英文名稱： 上皮細胞膜蛋白3抗體 0.2ml

Rhesus antibody Rh phospho-p107/RBL1(Ser975) 磷酸化視網(wǎng)膜母細胞瘤樣蛋白p107抗體 規(guī)格 0.1ml

Caspase-1 (ICE; CASP-1;P45) 天冬氨酸-胱氨酸特異性蛋白酶家族(抗原)Multi-class antibodies規(guī)格： 0.5mg

Anti-TIF1 beta/KAP1/FITC 熒光素標記轉(zhuǎn)錄中介因子Tif1β抗體IgGMulti-class antibodies規(guī)格： 0.2ml

NOS-3 (eNOS) endotheli cell NOS 合成酶-3(抗原)Multi-class antibodies規(guī)格： 0.5mg

微管相關(guān)蛋白抗體 Anti-Tau protein 0.1ml

phospho-STXBP1 (Ser515) 英文名稱： 磷酸化神經(jīng)突觸前膜胞內(nèi)蛋白抗體 0.1ml

phospho-Ephrin B (Ser300) 英文名稱： 磷酸化Ephrin B抗體 0.1ml

Rhesus antibody Rh phospho-LLGL1 + LLGL2(Ser650+Ser654) 磷酸化相似抑制基因HUGL抗體 規(guī)格 0.1ml

NOS-3 (eNOS) endotheli cell NOS 合成酶-3(抗原)Multi-class antibodies規(guī)格： 0.5mg

Mtp53(Mutant type p53) 突變型P53抗原Multi-class antibodies規(guī)格： 0.5mg

Anti-ASPP2/p53BP2 P53凋亡刺激蛋白2抗體Multi-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh phospho-AQP2(Ser261) 磷酸化水通道蛋白2抗體 規(guī)格 0.1ml

tau 濃縮液 0.1ml 進口分裝

Ube2L3 英文名稱： 泛素蛋白連接酶L3抗體 0.2ml

DHRS7 英文名稱： 視網(wǎng)膜短鏈脫氫酶/還原酶家族7抗體 0.2ml

Anti-ASPP2/p53BP2 P53凋亡刺激蛋白2抗體Multi-class antibodies規(guī)格： 0.2ml

IL21 Protein Rat 重組大鼠 IL21 / Ierleukin 21 蛋白 (His 標簽)

胰蛋白酶(含10mL 酶解緩沖液) 1mL

HA Others H5N1 甲型 H5N1 (A/Cambodia/S1211394/2008) 血凝素HA1 (Hemagglutinin) 人細胞裂解液 (陽性對照)

U-373MG-EGFP-puro[U-373MG-pLVT7](慢病毒構(gòu)建穩(wěn)定株)人 U-373MG-EGFP-puro[U-373MG-pLVT7] (leiviral consuct stable sain) of human brain glioma cells 1640+10%Hyclone 滅活血清+2μg/ml puromycin

CM-H020人胰島β細胞完全培養(yǎng)基100mL

人表皮角質(zhì)細胞-新生兒(HEK-n)( 5×105 ) 細胞名稱 種屬

CM-H092人脈絡(luò)膜細胞完全培養(yǎng)基100mL

KIR2DL1 Others Human 人 KIR2DL1 / CD158a 人細胞裂解液 (陽性對照)

小鼠晶狀體上皮細胞完全培養(yǎng)基 100mL

OPM-2-UBC-EGFP(穩(wěn)定株)人細胞 OPM-2-UBC-EGFP (stable sain) human myeloma cell 1640+20% FBS(熱滅活)

IL17F Protein Human 重組人 IL-17F / Ierleukin-17F 蛋白 (His 標簽)

人雪旺細胞 ( HSC) ( 5×105 ) 小鼠神經(jīng)干細胞 Mouse

凝血酶(凝血因子II)激活肽1抗體說明書 IL21 Protein Rat 重組大鼠 IL21 / Ierleukin 21 蛋白 (His 標簽)

胰蛋白酶(含10mL 酶解緩沖液) 1mL

HA Others H5N1 甲型 H5N1 (A/Cambodia/S1211394/2008) 血凝素HA1 (Hemagglutinin) 人細胞裂解液 (陽性對照)

U-373MG-EGFP-puro[U-373MG-pLVT7](慢病毒構(gòu)建穩(wěn)定株)人 U-373MG-EGFP-puro[U-373MG-pLVT7] (leiviral consuct stable sain) of human brain glioma cells 1640+10%Hyclone 滅活血清+2μg/ml puromycin

CM-H020人胰島β細胞完全培養(yǎng)基100mL

人表皮角質(zhì)細胞-新生兒(HEK-n)( 5×105 ) 細胞名稱 種屬