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抗體的生物素化標(biāo)記實(shí)驗(yàn)要點(diǎn)：

1. 磷酸化一氧化氮合成酶3（內(nèi)皮型）抗體規(guī)格 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-Phospho-eNOS (Thr495) 

中文名稱   磷酸化一氧化氮合成酶3（內(nèi)皮型）抗體規(guī)格 

別    名  eNOS (phospho T495); p-eNOS (phospho T495); eNOS (Phospho-Thr495); cNOS; eNOS; Constitutive NOS; EC NOS; ecNOS; Endothelial nitric oxidase synthase; Endothelial nitric oxide synthase; Endothelial nitric oxide synthase 3; Endothelial NOS; Nitric oxide synthase 3 (endothelial cell); Nitric oxide synthase 3; Nitric oxide synthase 3 endothelial cell; Nitric oxide synthase endothelial; nitric oxide synthase, endothelial; NOS 3; NOS III; NOS type III; NOS3; NOSIII; NOS3_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, Mouse, Rat, Dog, Pig, Cow, Rabbit, Sheep, Guinea Pig 

產(chǎn)品類型   一抗 磷酸化抗體  

研究領(lǐng)域    免疫學(xué) 神經(jīng)生物學(xué) 信號(hào)轉(zhuǎn)導(dǎo) 細(xì)胞凋亡 轉(zhuǎn)錄調(diào)節(jié)因子 激酶和磷酸酶

蛋白分子量  predicted molecular weight: 133kDa

性    狀   Lyophilized or Liquid

免 疫 原 KLH conjugated Synthesised phosphopeptide derived from human eNOS around the phosphorylation site of Thr495 

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

磷酸化一氧化氮合成酶3（內(nèi)皮型）抗體規(guī)格 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 eNOS is a calcium/calmodulin dependent enzyme which undergoes several post translational modifications, including acylation with myristate and palmitate, and phosphorylation on numerous residues. As with the other members of the NOS family, eNOS derives the diffusible multifunctional second messenger NO from L arginine through a series of reactions in which L citrulline is an intermediate. eNOS plays an important role in controlling vascular tone, platelet aggregation, and cardiac myocyte function.

Function : Produces nitric oxide (NO) which is implicated in vascular smooth muscle relaxation through a cGMP-mediated signal transduction pathway. NO mediates vascular endothelial growth factor (VEGF)-induced angiogenesis in coronary vessels and promotes blood clotting through the activation of platelets.Isoform eNOS13C: Lacks eNOS activity, dominant-negative form that may down-regulate eNOS activity by forming heterodimers with isoform 1.Subunit : Homodimer. Interacts with NOSIP and NOSTRIN.Subcellular Location : Cell membrane. Membrane, caveola. Cytoplasm, cytoskeleton. Golgi apparatus. Note=Specifically associates with actin cytoskeleton in the G2 phase of the cell cycle; which is favored by interaction with NOSIP and results in a reduced enzymatic activity.Tissue Specificity : Platelets, placenta, liver and kidney.Post-translational modifications : Phosphorylation by AMPK at Ser-1177 in the presence of Ca(2+)-calmodulin (CaM) activates activity. In absence of Ca(2+)-calmodulin, AMPK also phosphorylates Thr-495, resulting in inhibition of activity. Phosphorylation of Ser-114 by CDK5 reduces activity.Similarity : Belongs to the NOS family.Contains 1 FAD-binding FR-type domain.Contains 1 flavodoxin-like domain.Database links : UniProtKB/Swiss-Prot: P29474.3 humanUniProtKB/Swiss-Prot: Q62600.4 ratUniProtKB/Swiss-Prot: P70313.4 mouse催化生物體內(nèi)一氧化氮（NO）生成的酶。分神經(jīng)型一氧化氮合成的酶（nNOS or NOS-1）、誘導(dǎo)型一氧化氮合成的酶（iNOS or NOS-2）、內(nèi)皮型一氧化氮合成的酶（eNOS or NOS-3）。此抗體識(shí)別分子量為135kDa的內(nèi)皮型一氧化氮合成的酶。

抗體的鑒定：

1）磷酸化一氧化氮合成酶3（內(nèi)皮型）抗體規(guī)格 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?？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)率來表示。交叉反應(yīng)率可用競爭抑制試驗(yàn)測(cè)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(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。

Anti-Phospho-EphA2 (Tyr594) /FITC 熒光素標(biāo)記兔抗人、大、小鼠磷酸化酪氨酸蛋白激酶A2受體抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Anti-Phospho-MSK1 (Ser360)/FITC 熒光素標(biāo)記磷酸化有絲分裂原和應(yīng)激活化型蛋白激酶1抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh C1orf51 1號(hào)染色體開放閱讀框51抗體 規(guī)格 0.2ml

TmR2(GDNFR Beta;TRN-R;GFR-alpha-2;NTNR-2) 膠質(zhì)細(xì)胞系源性神經(jīng)營養(yǎng)因子(多肽) 0.5mg

HLC3 英文名稱： 癌基因蛋白3抗體 0.2ml

Rhesus antibody Rh RSRC2 蛋白抗體 規(guī)格 0.2ml

Anti-Phospho-MSK1 (Ser360)/FITC 熒光素標(biāo)記磷酸化有絲分裂原和應(yīng)激活化型蛋白激酶1抗體IgGMulti-class antibodies規(guī)格： 0.2ml

VWF (Von Willebrand Factor) 假性因子/性因子抗原Multi-class antibodies規(guī)格： 0.5mg

Anti-Caveolin-1 細(xì)胞質(zhì)膜微囊蛋白-1抗體Multi-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh PCGF1 表觀抑制因子Polycomb家族成員PCGFl蛋白抗體 規(guī)格 0.2ml

PRLR Kit Human 人 PRLR / Prolactin Receptor ELISA配對(duì)抗體 ELISA

TLR7 英文名稱： Toll樣受體7抗體 0.1ml

caspase-8 subunit p18 英文名稱： 半胱酸蛋白酶8抗體 0.2ml

Anti-Caveolin-1 細(xì)胞質(zhì)膜微囊蛋白-1抗體Multi-class antibodies規(guī)格： 0.2ml

Sema3A (semaphorin 3A) 臂板蛋白3A(多肽)Multi-class antibodies規(guī)格： 0.5mg

Anti-BECN1 BECLIN-1抗體Multi-class antibodies規(guī)格： 0.1ml

Rhesus antibody Rh PGT/Slco2a1 溶質(zhì)載體蛋白家族21成員2抗體 規(guī)格 0.2ml

Glutathione S-Transferase pi1:60 濃縮液 0.1ml 進(jìn)口分裝

UCHL5IP 英文名稱： 泛素羧基末端水解酶5互相作用蛋白1抗體 0.2ml

phospho-Caveolin-2(Tyr19) 英文名稱： 磷酸化細(xì)胞質(zhì)膜微囊蛋白-2抗體 0.1ml

Anti-BECN1 BECLIN-1抗體Multi-class antibodies規(guī)格： 0.1ml

TNFSF11 Protein Cynomolgus 重組食蟹猴 RANKL / OPGL / TNFSF11 蛋白 (Fc 標(biāo)簽)

caski

ERBB3 Others Rhesus 恒河猴 HER3 / ErbB3 人細(xì)胞裂解液 (陽性對(duì)照)

Hepa1-6-EGFP(CMV-GFP慢病毒構(gòu)建穩(wěn)定株)小鼠細(xì)胞 Hepa1-6-EGFP (CMV-GFP leiviral consuct stable sain) of hepatocellular carcinoma cells in mice DMEM+10% FBS

小鼠B淋巴細(xì)胞雜交瘤細(xì)胞;SH3

人主動(dòng)脈平滑肌細(xì)胞 (HASMC)( 5×105 ) HK-2, 人腎皮質(zhì)近曲小管上皮細(xì)胞 Human

CL-0462UMR-106(大鼠細(xì)胞)5×106cells/瓶×2

HA Others H7N9 甲型 H7N9 (A/Shanghai/2/2013) 血凝素 (Hemagglutinin / HA) 桿狀病毒-昆蟲細(xì)胞裂解液 (陽性對(duì)照)

人髓核細(xì)胞RNAHNPC miRNA5 μg

兔角膜前基質(zhì)層成纖維細(xì)胞;RCFBF 癌細(xì)胞，MCF7細(xì)胞 Cl.Ly1+2-/9細(xì)胞，小鼠T淋巴細(xì)胞

人臍內(nèi)皮細(xì)胞;HUV-EC-C

HA Others H5N1 甲型 H5N1 (A/goose/Guiyang/337/2006) 血凝素 (Hemagglutinin / HA) 人細(xì)胞裂解液 (陽性對(duì)照)

磷酸化一氧化氮合成酶3（內(nèi)皮型）抗體規(guī)格 TNFSF11 Protein Cynomolgus 重組食蟹猴 RANKL / OPGL / TNFSF11 蛋白 (Fc 標(biāo)簽)

caski

ERBB3 Others Rhesus 恒河猴 HER3 / ErbB3 人細(xì)胞裂解液 (陽性對(duì)照)

Hepa1-6-EGFP(CMV-GFP慢病毒構(gòu)建穩(wěn)定株)小鼠細(xì)胞 Hepa1-6-EGFP (CMV-GFP leiviral consuct stable sain) of hepatocellular carcinoma cells in mice DMEM+10% FBS

小鼠B淋巴細(xì)胞雜交瘤細(xì)胞;SH3

人主動(dòng)脈平滑肌細(xì)胞 (HASMC)( 5×105 ) HK-2, 人腎皮質(zhì)近曲小管上皮細(xì)胞 Human