抗体的生物素化标记实验要点：

1. 神经细胞分化因子1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*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。

产品订购信息：
英文名称  Anti-NeuroD1/NEUROD 

中文名称   神经细胞分化因子1抗体说明书 

别    名  atonal; basic helix loop helix transcription factor; bHLHa3; class A basic helix loop helix protein 3; Class A basic helix-loop-helix protein 3; MODY 6; MODY6; NDF1_HUMAN; NeuroD1; neurogenic helix loop helix protein NEUROD; Beta cell E box transactivator 2; BETA2; BHF 1; BHF1; NEUROD; Neurogenic differentiation 1; Neurogenic differentiation factor 1; NIDDM; BHLHA3; NEUROD1.

浓    度   1mg/1ml

规 格  0.1ml/100μg 0.2ml/200μg

抗体来源   Rabbit

克隆类型   polyclonal

交叉反应   Human, Mouse, Rat, Dog, Pig, Cow 

产品类型   一抗  

研究领域     心 免疫学 染色质和核信号 神经生物学 干细胞 细胞类型标志物 表观遗传学

蛋白分子量  predicted molecular weight: 40kDa

性    状   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human NeuroD1 C-terminus 

亚    型   IgG

纯化方法   affinity purified by Protein A

储 存 液   0.01M PBS, pH 7.4 with 10 mg/ml BSA and 0.1% Sodium azide

神经细胞分化因子1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=1:100-500

（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 NeuroD1 (Neurogenic Differentiation factor 1, also known as BETA2, beta-cell E-box transactivator 2) is a 40kDa protein, a member of the bHLH family. NeuroD1 is a homolog of the Drosophila atonal gene. NeuroD1 is widely expressed during development in the mammalian brain and pancreas where it acts as a transcriptional activator, and binds to insulin gene E-box. NeuroD1 has also been reported to be a differentiation factor in neurogenesis. Changes in NeuroD1 gene expression have been linked to diabetes mellitus type II and maturity-onset diabetes of the young type VI. NeuroD1 expression can be induced by glucose in beta cells.

Function : Acts as a transcriptional activator: mediatestranscriptional activation by binding to E box-containing promoterconsensus core sequences 5'-CANNTG-3'. Associates with the p300/CBPtranscription coactivator complex to stimulate transcription of thesecretin gene as well as the gene encoding the cyclin-dependentkinase inhibitor CDKN1A. Contributes to the regulation of severalcell differentiation pathways, like those that promote theformation of early retinal ganglion cells, inner ear sensoryneurons, granule cells forming either the cerebellum or the dentategyrus cell layer of the hippocampus, endocrine islet cells of thepancreas and enteroendocrine cells of the small intestine. Togetherwith PAX6 or SIX3, is required for the regulation of amacrine cellfate specification. Also required for dendrite morphogenesis andmaintenance in the cerebellar cortex. Associates with chromatin toenhancer regulatory elements in genes encoding key transcriptionalregulators of neurogenesis (By similarity).Subunit : Interacts (via helix-loop-helix motif domain) with EP300(via C-terminus) (By similarity). Heterodimer with TCF3/E47; theheterodimer is inhibited in presence of ID2, but not NR0B2, toE-box element. Efficient DNA-binding requires dimerization withanother bHLH protein. Interacts with RREB1. Interacts with EP300;the interaction is inhibited by NR0B2. Interacts with TCF3; theinteraction is inhibited by ID2.Subcellular Location : Cytoplasm (By similarity). Nucleus. Note=Inpancreatic islet cells, shuttles to the nucleus in response toglucose stimulation (By similarity). Colocalizes with NR0B2 in thenucleus.Post-translational modifications : Phosphorylated. In islet cells, phosphorylated on Ser-274upon glucose stimulation; which may be required for nuclearlocalization. In activated neurons, phosphorylated on Ser-335;which promotes dendritic growth. Phosphorylated by MAPK1;phosphorylation regulates heterodimerization and DNA-bindingactivities. Phosphorylation on Ser-266 and Ser-274 increasestransactivation on the insulin promoter in glucose-stimulatedinsulinoma cells (By similarity).DISEASE : Maturity-onset diabetes of the young 6 (MODY6)[MIM:606394]: A form of diabetes that is characterized by anautosomal dominant mode of inheritance, onset in childhood or earlyadulthood (usually before 25 years of age), a primary defect ininsulin secretion and frequent insulin-independence at thebeginning of the disease. Note=The disease is caused by mutationsaffecting the gene represented in this entry.Similarity : Contains 1 bHLH (basic helix-loop-helix) domain.Database links : UniProtKB/Swiss-Prot: Q13562.3神经生物学相关蛋白(Neurobiology)

抗体的鉴定：

1）神经细胞分化因子1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。

CPAF(Human chlamydial protease-like activity factor) ELISA Kit 人衣原体蛋白酶样活性因子Multi-class antibodies规格： 48T

Anti-Integrin alpha V/CD51 整合素αV抗体Multi-class antibodies规格： 0.2ml

Rhesus antibody Rh LDOC1/BCUR1 亮氨酸拉链下调因子1抗体 规格 0.2ml

i-PTH(Human intact Parathormone) ELISA Kit 人全段甲状旁腺素 96T

PML 英文名称： 早幼粒细胞蛋白抗体 0.2ml

Cadherin 10 英文名称： 钙粘附分子10抗体 0.2ml

Anti-Integrin alpha V/CD51 整合素αV抗体Multi-class antibodies规格： 0.2ml

MUC5B ELISA Kit 小鼠粘蛋白/粘液素5BMulti-class antibodies规格： 48T

Anti-BMP2/FITC 荧光素标记骨形态发生蛋白-2抗体IgGMulti-class antibodies规格： 0.2ml

Rhesus antibody Rh Ephrin B3 酪氨酸蛋白激酶B3抗体 规格 0.2ml

Dolichos bifows agglutinin,DBA ELISA Kit 双花扁豆凝集素 96T

MTP18 英文名称： 线粒体蛋白18抗体 0.2ml

ACF1 英文名称： 溴区结构域相邻锌指蛋白1A抗体 0.2ml

Anti-BMP2/FITC 荧光素标记骨形态发生蛋白-2抗体IgGMulti-class antibodies规格： 0.2ml

Anti-phospho-GCN2 (Thr667) /FITC 荧光素标记兔抗人、大、小鼠磷酸化蛋白激酶GCN2蛋白抗体IgGMulti-class antibodies规格： 0.2ml

Anti-N-ras/FITC 荧光素标记原癌基因抗体IgGMulti-class antibodies规格： 0.2ml

Rhesus antibody Rh BNP(H1G3) 人脑素单克隆抗体 规格 0.1ml

GTP-CH-1 三磷酸鸟苷环水解酶(抗原) 0.5mg

HEY2 英文名称： 转录因子HEY2蛋白抗体 0.1ml

Rhesus antibody Rh RelB 转录因子RelB蛋白抗体 规格 0.2ml

Anti-N-ras/FITC 荧光素标记原癌基因抗体IgGMulti-class antibodies规格： 0.2ml

BG250g用于蓝藻细菌培养

LB肉汤，Lennox 加富培养基，用于分子微生物研究中保存和培养重组大肠杆菌菌株 500g incubation media LB肉汤，Lennox 加富培养基，用于分子微生物研究中保存和培养重组大肠杆菌菌株 500g

调料葡萄球菌 产杆菌肽A 支/瓶

XLD培养基平板(9cm)选择性培养基，主要用于分离志贺氏菌属，亦可用于分离沙门氏菌

DRBC琼脂 250g 用于食品中霉菌及酵母菌分离培养

甘露醇发酵培养基250g生化试验培养基，用于细菌(如金黄色葡萄球菌)甘露醇发酵试验(GB标准)

煌绿乳糖胆盐肉汤(BGLB) 1000(g) incubation media 煌绿乳糖胆盐肉汤(BGLB) 1000(g)

LB肉汤 LB Broth 250克 用于分子生物学实验中大肠杆菌的保存和培养

40%尿素水5ml*尿素酶琼脂基础incubationmedia40%尿素水5ml*尿素酶琼脂基础

RVSBroth

神经细胞分化因子1抗体说明书 RS琼脂 250(g)  incubation media RS琼脂 250(g)

AHM鉴别用培养基 250(g)  incubation media AHM鉴别用培养基 250(g)

脱脂奶蔗糖胰蛋白胨琼脂培养基 250(g)  incubation media 脱脂奶蔗糖胰蛋白胨琼脂培养基 250(g)

蔗糖胰蛋白胨肉汤 250(g)  incubation media 蔗糖胰蛋白胨肉汤 250(g)