产品订购信息：
英文名称  Anti-UBE2D3 

中文名称   泛素蛋白连接酶D3抗体说明书 

别    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.

浓    度   1mg/1ml

规 格  0.2ml/200μg

抗体来源   Rabbit

克隆类型   polyclonal

交叉反应   Human, Mouse, Rat, Chicken, Pig, Cow, Horse, Rabbit, Zebrafish 

产品类型   一抗  

研究领域     细胞生物 免疫学 信号转导 表观遗传学

蛋白分子量  predicted molecular weight: 16 kDa

性    状   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human UBE2D3 

亚    型   IgG

纯化方法   affinity purified by Protein A

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

泛素蛋白连接酶D3抗体说明书 产品应用    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

（石蜡切片需做抗原修复） 

 not yet tested in other applications.

 optimal dilutions/concentrations should be determined by the end user.  

保存条件  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. 

产品介绍 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 大鼠基质细胞衍生因子1aMulti-class antibodies规格： 48T

Anti-Phospho-Tuberin/TSC2 (Ty*0) 磷酸化马铃薯球蛋白(性硬化)抗体Multi-class antibodies规格： 0.1ml

Rhesus antibody Rh GDIA1 转移抑制基因GDIA1抗体 规格 0.1ml

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

eNOS 英文名称： 合成酶-3(内皮型)抗体 0.1ml

ADAMTS13 英文名称： 整合素样金属蛋白酶与凝血酶13型抗体 0.2ml

Anti-Phospho-Tuberin/TSC2 (Ty*0) 磷酸化马铃薯球蛋白(性硬化)抗体Multi-class antibodies规格： 0.1ml

Anti-SMMHC/MYH11/FITC 荧光素标记平滑肌肌球蛋白重链抗体IgGMulti-class antibodies规格： 0.2ml

Anti-Phospho-Jak1 (Tyr1022/1023) /FITC 荧光素标记磷酸化蛋白质酪氨酸激酶JAK-1抗体IgGMulti-class antibodies规格： 0.2ml

Rhesus antibody Rh CCDC69 卷曲螺旋结构域蛋白69抗体 规格 0.2ml

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

HPV18 E6 protein 英文名称： 人类状瘤病毒18 E6单克隆抗体 0.2ml

Rhesus antibody Rh SNTG2/Syntrophin 5 肌蛋白结合蛋白γ2抗体 规格 0.2ml

Anti-Phospho-Jak1 (Tyr1022/1023) /FITC 荧光素标记磷酸化蛋白质酪氨酸激酶JAK-1抗体IgGMulti-class antibodies规格： 0.2ml

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

Anti-LTB4/Leukotriene B4 白三烯B4抗体Multi-class antibodies规格： 0.2ml

Rhesus antibody Rh KiSS1R/GPR54 G蛋白偶联受体54抗体 规格 0.1ml

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

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

Phospho-cdc2* (Ser124) 英文名称： 磷酸化细胞分裂周期蛋白25抗体 0.1ml

Anti-LTB4/Leukotriene B4 白三烯B4抗体Multi-class antibodies规格： 0.2ml

PDHA1 Others Human 人 PDHA1 / C54G1 (aa 30-390) 杆状病毒-昆虫细胞裂解液 (阳性对照)

CM-H047人肝窦内皮细胞完全培养基100mL

人少突胶质细胞HO

非洲绿猴肾细胞系/HCV-NS3;Vero-HCV-NS3 人细胞，A-704细胞 AGS(胃腺癌细胞)

人神经母细胞瘤细胞;SH-SY5Y

FGF18 Others Mouse 小鼠 FGF18 / FGF-18 人细胞裂解液 (阳性对照)

CL-0386MPC-11(小鼠浆细胞瘤)5×106cells/瓶×2

EFNB1 Others Rat 大鼠 Ephrin-B1 / EFNB1 人细胞裂解液 (阳性对照)

小鼠气管平滑肌细胞完全培养基 100mL

EJ人细胞 Human bladder cancer cell line EJ 1640+10% FBS

IL36B Protein Mouse 重组小鼠 IL1F8 / IL36b 蛋白

NCI-H23(人非小细胞细胞) 5×106cells/瓶×2 THP-1(单核细胞)

泛素蛋白连接酶D3抗体说明书 CL-0383MDA-MB-436(人癌细胞)5×106cells/瓶×2

CXADR Others Mouse 小鼠 CXADR / CAR 人细胞裂解液 (阳性对照)

人角膜上皮细胞总RNAHCEpiC NA

ZR75-1细胞，人癌细胞 129小鼠ES细胞,D3细胞 WM35, 人细胞

MV3(人细胞) 5×106cells/瓶×2

HUAEC Pellet 人脐动脉内皮细胞团块 > 1 mio.cells 神经细胞培养基NM-prf

抗体的生物素化标记实验要点：

1. 泛素蛋白连接酶D3抗体说明书 如在反应混合液中有叠氮钠或游离氨基存在,会抑制标记反应。因此,蛋白质在反应前要对 0.1mol/L碳酸氢钠缓冲液或0.5mol/L硼酸缓冲液充分透析；

2.所用的NHSB及待生物素化蛋白质之间的分子比按蛋白质表面的ε-氨基的密度会有所不同,选择不当则影响标记的效率,应先用几个不同的分子比来筛选最适条件；

3.用NHSB量过量也是不利的,抗原的结合位点可能因此被封闭,导致抗体失活；

4.由于抗体的氨基不易接近可能造成生物素化不足,此时可加入去污剂如 Triton x-100, Tween20等；

5.当游离ε-氨基(赖氨酸残基的氨基)存在于抗体的抗原结合位点时,或位于酶的催化位点时,生物素化会降低或损伤抗体蛋白的结合力或活性；

6.生物素还可能与不同的功能基团,如羰基、氨基、巯基、异咪唑基及苯酚基,也可与糖基共价结合；

7.交联反应后,应充分透析,否则,残余的生物素会对生物素化抗体与亲和素的结合产生竞争作用；

8.在细胞的荧光标记实验中,中和亲和素的本底低,但由于链霉亲和素含有少量正电荷,故对某些细胞可导致高本底。

抗体的鉴定：

1）泛素蛋白连接酶D3抗体说明书 抗体的效价鉴定：不管是用于诊断还是用于,制备抗体的目的都是要求较高效价。不同的抗原制备的抗体,要求的效价不一。鉴定效价的方法很多,包括有试管凝集反应,琼脂扩散试验,酶联免疫吸附试验等。常用的抗原所制备的抗体一般都有约成的鉴定效价的方法,以资比较。如制备抗抗体的效价,一般就采用琼脂扩散试验来鉴定。

2）抗体的特异性鉴定：抗体的特异性是指与相应抗原或近似抗原物质的识别能力。抗体的特异性高,它的识别能力就强。衡量特异性通常以交叉反应率来表示。交叉反应率可用竞争抑制试验测定。以不同浓度抗原和近似抗原分别做竞争抑制曲线,计算各自的结合率,求出各自在IC50时的浓度,并按公式计算交叉反应率。 

如果所用抗原浓度IC50浓度为pg/管,而一些近似抗原物质的IC50浓度几乎是无穷大时,表示这一抗血清与其他抗原物质的交叉反应率近似为0,即该血清的特异性较好。

3）抗体亲和力：是指抗体和抗原结合的牢固程度。亲和力的高低是由抗原分子的大小,抗体分子的结合位点与抗原决定簇之间立体构型的合适度决定的。有助于维持抗原抗体复合物稳定的分子间力有氢键,疏水键,侧链相反电荷基因的库仑力,范德华力和空间斥力。亲和力常以亲和常数K表示,K的单位是L/mol。抗体亲和力的测定对抗体的筛选,确定抗体的用途,验证抗体的均一性等均有重要意义。