抗體的生物素化標(biāo)記實(shí)驗(yàn)要點(diǎn)：

1. 泛素蛋白連接酶E1抗體說(shuō)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(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(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-UBCH6/UBE2E1 

中文名稱   泛素蛋白連接酶E1抗體說(shuō)明書 

別    名  UB2E1_HUMAN; UbcH6; UBE2E1; Ubiquitin carrier protein E1; Ubiquitin-conjugating enzyme E2 E1; Ubiquitin-protein ligase E1.

濃    度   1mg/1ml

規(guī) 格  0.2ml/200μg

抗體來(lái)源   Rabbit

克隆類型   polyclonal

交叉反應(yīng)   Mouse, Rat, Chicken, Pig, Cow, Sheep 

產(chǎn)品類型   一抗  

研究領(lǐng)域    

蛋白分子量  predicted molecular weight: 21kDa

性    狀   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human UBCH6/UBE2E1 

亞    型   IgG

純化方法   affinity purified by Protein A

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

泛素蛋白連接酶E1抗體說(shuō)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=1:50-200

（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 ubiquitin (Ub) pathway involves three sequential enzymatic steps that facilitate the conjugation of Ub and Ub-like molecules to specific protein substrates. The first step requires the ATP-dependent activation of the Ub C-terminus and the assembly of multi-Ub chains by the Ub-activating enzyme known as the E1 component. The Ub chain is then conjugated to the Ub-conjugating enzyme (E2) to generate an intermediate Ub-E2 complex. The Ub-ligase (E3) then catalyzes the transfer of Ub from E2 to the appropriate protein substrate. UBE2E1 and UBE2L3, also designated UBCH6 and UBCH7 respectively in human, are E2 conjugating enzymes that interact with various proteins. Specifically, UBE2E1 interacts with the tumor suppressor protein TSSC5. UBE2L3 has been shown to mediate c-fos degradation, NF-kB maturation, human papilloma virus-mediated p53 and Myc protein degradation.

Function : Accepts ubiquitin from the E1 complex and catalyzes its covalent attachment to other proteins. Catalyzes the covalent attachment of ISG15 to other proteins. Mediates the selective degradation of short-lived and abnormal proteins. In vitro also catalyzes 'Lys-48'-linked polyubiquitination.

Subunit : Interacts with RNF14.

Subcellular Location : Nucleus.

Post-translational modifications : ISGylation suppresses ubiquitin E2 enzyme activity.

Similarity : Belongs to the ubiquitin-conjugating enzyme family.

Database links : UniProtKB/Swiss-Prot: P51965.1

抗體的鑒定：

1）泛素蛋白連接酶E1抗體說(shuō)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(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。

Anti-HSP-27/FITC 熒光素標(biāo)記HSP-27蛋白抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Anti-Pin1/FITC 熒光素標(biāo)記肽基脯氨酞順?lè)串悩?gòu)酶Pinl抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh ASGPR1/ASGR1 唾液酸糖蛋白受體1抗體 規(guī)格 0.1ml

Donkey Anti-Goat IgG/PE-Cy7 PE-Cy7標(biāo)記的驢抗羊IgG 0.1ml

GLT6D1 英文名稱： 糖基轉(zhuǎn)移酶6結(jié)構(gòu)1抗體 0.2ml

Rhesus antibody Rh Rabbit Anti-mouse IgG/Cy5.5 Cy5.5標(biāo)記的兔抗小鼠IgG 規(guī)格 0.1ml

Anti-Pin1/FITC 熒光素標(biāo)記肽基脯氨酞順?lè)串悩?gòu)酶Pinl抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Anti-Phospho-Gab1 (Tyr627) /FITC 熒光素標(biāo)記兔抗人、大、小鼠磷酸化接頭蛋白Gab 1抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Anti-NOS-2/iNOS /FITC 熒光素標(biāo)記合成酶-2抗體(誘導(dǎo)型)IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh BRMS-1 癌轉(zhuǎn)移抑制基因1抗體 規(guī)格 0.1ml

Leptin 瘦素(多肽抗原) 0.5mg

HOXA11 英文名稱： 同源盒蛋白HOXA11抗體 0.1ml

Rhesus antibody Rh RhoA RhoA抗體 規(guī)格 0.1ml

Anti-NOS-2/iNOS /FITC 熒光素標(biāo)記合成酶-2抗體(誘導(dǎo)型)IgGMulti-class antibodies規(guī)格： 0.2ml

RLN(Human Relaxin) ELISA Kit 人松弛肽/松弛素Multi-class antibodies規(guī)格： 48T

Anti-LDL 抗體Multi-class antibodies規(guī)格： 0.1ml

Rhesus antibody Rh KNTC1/Kinetochore 著絲點(diǎn)關(guān)聯(lián)蛋白1抗體 規(guī)格 0.2ml

MS(Human melatonin sulfate) ELISA Kit 人褪黑色素 96T

Phospho-PSD95 (Tyr236 + Tyr240) 英文名稱： 磷酸化突觸后密度蛋白95抗體 0.1ml

Calcipressin 3 英文名稱： 鈣調(diào)神經(jīng)磷酸酶調(diào)節(jié)蛋白3抗體(唐氏綜合征候選區(qū)域蛋白1樣蛋白3) 0.1ml

Anti-LDL 抗體Multi-class antibodies規(guī)格： 0.1ml

PROM1 Others Rat 大鼠 CD133 / PROM1 / Prominin 1 人細(xì)胞裂解液 (陽(yáng)性對(duì)照)

人少突膠質(zhì)前體細(xì)胞-懸浮生長(zhǎng)RNAHOPC-os miRNA5 μg

兔角膜前基質(zhì)層成纖維細(xì)胞;RCFBF

HRT-S1細(xì)胞，人細(xì)胞 人橫紋肌細(xì)胞,TE671 subline No.2細(xì)胞 小鼠源細(xì)胞

HL-7702(人肝正常細(xì)胞) 5×106cells/瓶×2

BACE1 Others Human 人 BACE1 / ASP2 人細(xì)胞裂解液 (陽(yáng)性對(duì)照)

CL-0451SW-13(人腎上腺皮質(zhì)癌細(xì)胞)5×106cells/瓶×2

CTSS Others Mouse 小鼠 Cathepsin S / CTSS 人細(xì)胞裂解液 (陽(yáng)性對(duì)照)

大鼠角膜上皮細(xì)胞完全培養(yǎng)基 100mL

Hep3B人細(xì)胞 Hep3B human hepatocarcinoma cells DMEM+10% FBS

IL18 Protein Human 重組人 IL18 / Ierleukin 18 / IGIF 蛋白 (GST 標(biāo)簽)

NCI-H520(人肺鱗癌細(xì)胞) 5×106cells/瓶×2 A549 [A-549](細(xì)胞)

泛素蛋白連接酶E1抗體說(shuō)明書 CL-0448SW 1990(人*癌細(xì)胞)5×106cells/瓶×2

VSIG4 Others Human 人 VSIG4 人細(xì)胞裂解液 (陽(yáng)性對(duì)照)

人骨骼肌成肌細(xì)胞RNAHSkMM miRNA5 μg

ScaBER細(xì)胞，人膀胱鱗癌細(xì)胞 小鼠細(xì)胞,C26細(xì)胞 RLFs, 大鼠

L-O2(人正常肝細(xì)胞) 5×106cells/瓶×2

NHEK.f single donor Pellet 正常人表皮角質(zhì)細(xì)胞團(tuán)塊(少年者) > 1 mio.cells 小鼠顆粒細(xì)胞MGC