抗体的生物素化标记实验要点：

1. 转录增强因子TEF3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。

产品订购信息：
英文名称  Anti-TEF3/TEAD4 

中文名称   转录增强因子TEF3抗体说明书 

别    名  EFTR 2; EFTR2; hRTEF 1B; hRTEF1B; MGC9014; Related to TEF 1; Related to TEF1; Related transcription enhancer factor 1B; RTEF1; TCF13L1; TEA domain family member 4; TEAD 4; TEAD-4; TEAD4; TEAD4_HUMAN; TEF 3; TEF-3; TEF3; TEFR 1; TEFR1; Transcription factor 13 (SV40 transcriptional enhancer factor) like 1; Transcription factor 13 like 1; Transcription factor 13-like 1; Transcription factor RTEF 1; Transcription factor RTEF-1; Transcription factor RTEF1; Transcriptional enhancer factor 1 related; Transcriptional enhancer factor 3; Transcriptional enhancer factor TEF 3; Transcriptional enhancer factor TEF-3; Transcriptional enhancer factor TEF3.

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, Dog, Pig, Cow, Rabbit, Zebrafish 

产品类型   一抗  

研究领域    心 转录调节因子 表观遗传学

蛋白分子量  predicted molecular weight: 48kDa

性    状   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human TEF3/TEAD4 

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

转录增强因子TEF3抗体说明书 产品应用    WB=1:100-500 ELISA=1:500-1000 IHC-P=1:100-500 IHC-F=1:100-500 ICC=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 TEF-3, is a 427 amino acid member of the transcriptional enhancer factor (TEF) family of proteins that are characterized by the presence of a TEA DNA-binding domain. Localized to the nucleus and expressed primarily in skeletal muscle, TEF-3 functions as a transcriptional regulator by binding specifically and non-cooperatively to the M-CAT motif found in the promotors of muscle-specific genes, thereby directing their subsequent expression. TEF-3 contains one TEA DNA-binding domain and is expressed as multiple isoforms due to alternative splicing events.

Function : ranscription factor which plays a key role in the Hippo signaling pathway, a pathway involved in organ size control and tumor suppression by restricting proliferation and promoting apoptosis. The core of this pathway is composed of a kinase cascade wherein MST1/MST2, in complex with its regulatory protein SAV1, phosphorylates and activates LATS1/2 in complex with its regulatory protein MOB1, which in turn phosphorylates and inactivates YAP1 oncoprotein and WWTR1/TAZ. Acts by mediating gene expression of YAP1 and WWTR1/TAZ, thereby regulating cell proliferation, migration and epithelial mesenchymal transition (EMT) induction. Binds specifically and non-cooperatively to the Sph and GT-IIC 'enhansons' (5'-GTGGAATGT-3') and activates transcription. Binds to the M-CAT motif.

Subunit : Interacts with YAP1 and WWTR1/TAZ.

Subcellular Location : Nucleus.

Tissue Specificity : Preferentially expressed in skeletal muscle. Lower levels in pancreas, placenta, and heart.

Similarity : Contains 1 TEA DNA-binding domain.

Database links : UniProtKB/Swiss-Prot: Q15561.3

抗体的鉴定：

1）转录增强因子TEF3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。

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