產(chǎn)品訂購(gòu)信息：
英文名稱  Anti-UBE2D3 

中文名稱   泛素蛋白連接酶D3抗體說(shuō)明書(shū) 

別    名  E2(17)KB3; MGC43926; MGC5416; UB2D3_HUMAN; UBC 4/5; UBC4/5; UBC4/5 homolog yeast; UBCH 5C; UBCH5C; Ube2d3; Ubiquitin carrier protein; Ubiquitin carrier protein D3; Ubiquitin conjugating enzyme E2 17 kDa 3; Ubiquitin conjugating enzyme E2 D3; Ubiquitin conjugating enzyme E2D 3 (homologous to yeast UBC4/5); Ubiquitin conjugating enzyme E2D 3 (UBC4/5 homolog yeast); Ubiquitin conjugating enzyme E2D 3; Ubiquitin protein ligase D3; Ubiquitin-conjugating enzyme E2 D3; Ubiquitin-conjugating enzyme E2(17)KB 3; Ubiquitin-conjugating enzyme E2-17 kDa 3; Ubiquitin-protein ligase D3.

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)   Human, Mouse, Rat, Chicken, Pig, Cow, Horse, Rabbit, Zebrafish 

產(chǎn)品類型   一抗  

研究領(lǐng)域     細(xì)胞生物 免疫學(xué) 信號(hào)轉(zhuǎn)導(dǎo) 表觀遺傳學(xué)

蛋白分子量  predicted molecular weight: 16 kDa

性    狀   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human UBE2D3 

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D3抗體說(shuō)明書(shū) 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 Accepts ubiquitin from the E1 complex and catalyzes its covalent attachment to other proteins. In vitro catalyzes 'Lys-11'-, as well as 'Lys-48'-linked polyubiquitination. Cooperates with the E2 CDC34 and the SCF(FBXW11) E3 ligase complex for the polyubiquitination of NFKBIA leading to its subsequent proteasomal degradation. Acts as an initiator E2, priming the phosphorylated NFKBIA target at positions 'Lys-21' and/or 'Lys-22' with a monoubiquitin. Ubiquitin chain elongation is then performed by CDC34, building ubiquitin chains from the UBE2D3-primed NFKBIA-linked ubiquitin. Acts also as an initiator E2, in conjunction with RNF8, for the priming of PCNA. Monoubiquitination of PCNA, and its subsequent polyubiquitination, are essential events in the operation of the DNA damage tolerance (DDT) pathway that is activated after DNA damage caused by UV or chemical agents during S-phase. Associates with the BRCA1/BARD1 E3 ligase complex to perform ubiquitination at DNA damage sites following ionizing radiation leading to DNA repair. Targets DAPK3 for ubiquitination which influences promyelocytic leukemia protein nuclear body (PML-NB) formation in the nucleus. In conjunction with the MDM2 and TOPORS E3 ligases, functions ubiquitination of p53/TP53. Supports NRDP1-mediated ubiquitination and degradation of ERBB3 and of BRUCE which triggers apoptosis. In conjunction with the CBL E3 ligase, targets EGFR for polyubiquitination at the plasma membrane as well as during its internalization and transport on endosomes. In conjunction with the STUB1 E3 quality control E3 ligase, ubiquitinates unfolded proteins to catalyze their immediate destruction.

Function : Accepts ubiquitin from the E1 complex and catalyzes its covalent attachment to other proteins. In vitro catalyzes 'Lys-11'-, as well as 'Lys-48'-linked polyubiquitination. Cooperates with the E2 CDC34 and the SCF(FBXW11) E3 ligase complex for the polyubiquitination of NFKBIA leading to its subsequent proteasomal degradation. Acts as an initiator E2, priming the phosphorylated NFKBIA target at positions 'Lys-21' and/or 'Lys-22' with a monoubiquitin. Ubiquitin chain elongation is then performed by CDC34, building ubiquitin chains from the UBE2D3-primed NFKBIA-linked ubiquitin. Acts also as an initiator E2, in conjunction with RNF8, for the priming of PCNA. Monoubiquitination of PCNA, and its subsequent polyubiquitination, are essential events in the operation of the DNA damage tolerance (DDT) pathway that is activated after DNA damage caused by UV or chemical agents during S-phase. Associates with the BRCA1/BARD1 E3 ligase complex to perform ubiquitination at DNA damage sites following ionizing radiation leading to DNA repair. Targets DAPK3 for ubiquitination which influences promyelocytic leukemia protein nuclear body (PML-NB) formation in the nucleus. In conjunction with the MDM2 and TOPORS E3 ligases, functions ubiquitination of p53/TP53. Supports NRDP1-mediated ubiquitination and degradation of ERBB3 and of BRUCE which triggers apoptosis. In conjunction with the CBL E3 ligase, targets EGFR for polyubiquitination at the plasma membrane as well as during its internalization and transport on endosomes. In conjunction with the STUB1 E3 quality control E3 ligase, ubiquitinates unfolded proteins to catalyze their immediate destruction (By similarity).

Subcellular Location : Cell membrane; Peripheral membrane protein. Endosome membrane; Peripheral membrane protein.

Similarity : Belongs to the ubiquitin-conjugating enzyme family.

SDF-1a/CXCL12 ELISA Kit 大鼠基質(zhì)細(xì)胞衍生因子1aMulti-class antibodies規(guī)格： 48T

Anti-Phospho-Tuberin/TSC2 (Ty*0) 磷酸化馬鈴薯球蛋白(性硬化)抗體Multi-class antibodies規(guī)格： 0.1ml

Rhesus antibody Rh GDIA1 轉(zhuǎn)移抑制基因GDIA1抗體 規(guī)格 0.1ml

TNF- Beta(Mouse Tumor necrosis factor Beta) ELISA KIT 小鼠壞死因子β 96T

eNOS 英文名稱： 合成酶-3(內(nèi)皮型)抗體 0.1ml

ADAMTS13 英文名稱： 整合素樣金屬蛋白酶與凝血酶13型抗體 0.2ml

Anti-Phospho-Tuberin/TSC2 (Ty*0) 磷酸化馬鈴薯球蛋白(性硬化)抗體Multi-class antibodies規(guī)格： 0.1ml

Anti-SMMHC/MYH11/FITC 熒光素標(biāo)記平滑肌肌球蛋白重鏈抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Anti-Phospho-Jak1 (Tyr1022/1023) /FITC 熒光素標(biāo)記磷酸化蛋白質(zhì)酪氨酸激酶JAK-1抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh CCDC69 卷曲螺旋結(jié)構(gòu)域蛋白69抗體 規(guī)格 0.2ml

Anti-HMGA1 (high mobility group AT-hook 1) 高遷移率族蛋白A1抗原 0.5mg

HPV18 E6 protein 英文名稱： 人類狀瘤病毒18 E6單克隆抗體 0.2ml

Rhesus antibody Rh SNTG2/Syntrophin 5 肌蛋白結(jié)合蛋白γ2抗體 規(guī)格 0.2ml

Anti-Phospho-Jak1 (Tyr1022/1023) /FITC 熒光素標(biāo)記磷酸化蛋白質(zhì)酪氨酸激酶JAK-1抗體IgGMulti-class antibodies規(guī)格： 0.2ml

E2(Human Estradiol) ELISA Kit 人雌二醇Multi-class antibodies規(guī)格： 48T

Anti-LTB4/Leukotriene B4 白三烯B4抗體Multi-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh KiSS1R/GPR54 G蛋白偶聯(lián)受體54抗體 規(guī)格 0.1ml

HDV IgG(Human Hepatitis D virus IgG) ELISA Kit 人IgG 96T

phospho-PLB(Ser16) 英文名稱： 磷酸化磷蛋白抗體 0.1ml

Phospho-cdc2* (Ser124) 英文名稱： 磷酸化細(xì)胞分裂周期蛋白25抗體 0.1ml

Anti-LTB4/Leukotriene B4 白三烯B4抗體Multi-class antibodies規(guī)格： 0.2ml

PDHA1 Others Human 人 PDHA1 / C54G1 (aa 30-390) 桿狀病毒-昆蟲(chóng)細(xì)胞裂解液 (陽(yáng)性對(duì)照)

CM-H047人肝竇內(nèi)皮細(xì)胞完全培養(yǎng)基100mL

人少突膠質(zhì)細(xì)胞HO

非洲綠猴腎細(xì)胞系/HCV-NS3;Vero-HCV-NS3 人細(xì)胞，A-704細(xì)胞 AGS(胃腺癌細(xì)胞)

人神經(jīng)母細(xì)胞瘤細(xì)胞;SH-SY5Y

FGF18 Others Mouse 小鼠 FGF18 / FGF-18 人細(xì)胞裂解液 (陽(yáng)性對(duì)照)

CL-0386MPC-11(小鼠漿細(xì)胞瘤)5×106cells/瓶×2

EFNB1 Others Rat 大鼠 Ephrin-B1 / EFNB1 人細(xì)胞裂解液 (陽(yáng)性對(duì)照)

小鼠氣管平滑肌細(xì)胞完全培養(yǎng)基 100mL

EJ人細(xì)胞 Human bladder cancer cell line EJ 1640+10% FBS

IL36B Protein Mouse 重組小鼠 IL1F8 / IL36b 蛋白

NCI-H23(人非小細(xì)胞細(xì)胞) 5×106cells/瓶×2 THP-1(單核細(xì)胞)

泛素蛋白連接酶D3抗體說(shuō)明書(shū) CL-0383MDA-MB-436(人癌細(xì)胞)5×106cells/瓶×2

CXADR Others Mouse 小鼠 CXADR / CAR 人細(xì)胞裂解液 (陽(yáng)性對(duì)照)

人角膜上皮細(xì)胞總RNAHCEpiC NA

ZR75-1細(xì)胞，人癌細(xì)胞 129小鼠ES細(xì)胞,D3細(xì)胞 WM35, 人細(xì)胞

MV3(人細(xì)胞) 5×106cells/瓶×2

HUAEC Pellet 人臍動(dòng)脈內(nèi)皮細(xì)胞團(tuán)塊 > 1 mio.cells 神經(jīng)細(xì)胞培養(yǎng)基NM-prf

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D3抗體說(shuō)明書(shū) 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。

抗體的鑒定：

1）泛素蛋白連接酶D3抗體說(shuō)明書(shū) 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。