抗体的生物素化标记实验要点：

1. 孕激素受体抗体说明书 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-PR (progesterone receptor) 

中文名称   孕激素受体抗体说明书 

别    名  NR3C3; Nuclear receptor subfamily 3 group C member 3; PGR; PR; PRA; PRB; Progesterone receptor; Progestin receptor form A; Progestin receptor form B.

浓    度   1mg/1ml

规 格  0.1ml/100μg 0.2ml/200μg 1ml/1mg

抗体来源   Rabbit

克隆类型   polyclonal

交叉反应   Human, Mouse, Rat, Dog, Horse, Rabbit 

产品类型   一抗  

研究领域     免疫学 染色质和核信号 信号转导 生长因子和激素 细胞膜受体 细胞生物标志物

蛋白分子量  predicted molecular weight: 103kDa

性    状   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human progesterone receptor 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

孕激素受体抗体说明书 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 Estrogen and progesterone receptor are members of a family of transcription factors that are regulated by the binding of their cognate ligands. The interaction of hormone-bound estrogen receptors with estrogen responsive elements(EREs) alters transcription of ERE-containing genes. The carboxy terminal region of the estrgen receptor contains the ligand binding domain, the amino terminus serves as the transactivation domain, and the DNA binding domain is centrally located. Two forms of estrogen receptor have been identified, ER alpha and ER beta. ER alpha and ER beta have been shown to be differentially activated by various ligands. The biological response to progesterone is mediated by two distinct forms of the human progesterone receptor (hPR-Aand hPR-B), which arise from alternative splicing. In most cells, hPR-B functions as a transcriptional activator of progesterone-responsive gene, whereas hPR-A function as a transcriptional inhibitor of all steroid hormone receptors.

Function : The steroid hormones and their receptors are involved in the regulation of eukaryotic gene expression and affect cellular proliferation and differentiation in target tissues. Progesterone receptor isoform B (PRB) is involved activation of c-SRC/MAPK signaling on hormone stimulation.

Isoform A is inactive in stimulating c-Src/MAPK signaling on hormone stimulation.

Subunit : Interacts with SMARD1 and UNC45A. Interacts with CUEDC2; the interaction promotes ubiquitination, decreases sumoylation, and repesses transcriptional activity. Interacts with PIAS3; the interaction promotes sumoylation of PR in a hormone-dependent manner, inhibits DNA-binding, and alters nuclear export. Interacts with SP1; the interaction requires ligand-induced phosphorylation on Ser-345 by ERK1/2 MAPK. Interacts with PRMT2.

Post-translational modifications : Phosphorylated on multiple serine sites. Several of these sites are hormone-dependent. Phosphorylation on Ser-294 occurs preferentially on isoform B, is highly hormone-dependent and modulates ubiquitination and sumoylation on Lys-388. Phosphorylation on Ser-102 and Ser-345 also requires induction by hormone. Basal phosphorylation on Ser-81, Ser-162, Ser-190 and Ser-400 is increased in response to progesterone and can be phosphorylated in vitro by the CDK2-A1 complex. Increased levels of phosphorylation on Ser-400 also in the presence of EGF, heregulin, IGF, PMA and FBS. Phosphorylation at this site by CDK2 is ligand-independent, and increases nuclear translocation and transcriptional activity. Phosphorylation at Ser-162 and Ser-294, but not at Ser-190, is impaired during the G(2)/M phase of the cell cycle. Phosphorylation on Ser-345 by ERK1/2 MAPK is required for interaction with SP1.

Sumoylation is hormone-dependent and represses transcriptional activity. Sumoylation on all three sites is enhanced by PIAS3. Desumoylated by SENP1. Sumoylation on Lys-388, the main site of sumoylation, is repressed by ubiquitination on the same site, and modulated by phosphorylation at Ser-294.

Similarity : Belongs to the nuclear hormone receptor family. NR3 subfamily.

Contains 1 nuclear receptor DNA-binding domain.

类固醇受体（Steroid Receptors）孕激素受体是一类位于孕酮靶组织细胞内或细胞表面的特异蛋白质，特异地与孕酮结合，所形成的细胞溶质孕酮-受体复合物随后与细胞核内的DNA结合，以启动蛋白质生物合成。孕酮受体有A和B两种，受雌激素诱导，半寿期很短。

抗体的鉴定：

1）孕激素受体抗体说明书 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。

Anti-VE-cadherin/CD/VECD/FITC 荧光素标记上皮型钙粘附分子/内皮钙粘蛋白抗体IgGMulti-class antibodies规格： 0.2ml

Anti-IL-2R Gamma/FITC 荧光素标记白介素2受体γ链抗体IgGMulti-class antibodies规格： 0.2ml

Rhesus antibody Rh CD337/NKp30/NCR3 细胞毒性受体NK-p30抗体 规格 0.1ml

Neurocan 神经粘蛋白抗原 0.5mg

ICAM5 英文名称： 细胞间粘附分子5抗体 0.1ml

Rhesus antibody Rh SUMO 1 类泛素蛋白抗体 规格 0.1ml

Anti-IL-2R Gamma/FITC 荧光素标记白介素2受体γ链抗体IgGMulti-class antibodies规格： 0.2ml

Human retinoic acid inducible gene/RIG-like receptor,RLR-9 ELISA Kit 人视黄酸诱导基因/RIG-1样受体Multi-class antibodies规格： 48T

Anti-Phospho-Smad1 (Ser206) 磷酸化细胞信号转导分子Smad-1抗体Multi-class antibodies规格： 0.1ml

Rhesus antibody Rh Goat Anti-rabbit IgG/Cy7 Cy7标记的羊抗兔IgG 规格 0.1ml

AQP-3(Mouse Aquaporin 3) ELISA Kit 小鼠水通道蛋白3 96T

Natriuretic Peptide Receptor A 英文名称： 利肽受体A抗体 0.2ml

bFGF 英文名称： 碱性成纤维细胞生长因子抗体 0.1ml

Anti-Phospho-Smad1 (Ser206) 磷酸化细胞信号转导分子Smad-1抗体Multi-class antibodies规格： 0.1ml

SDH(Human sorbitol dehydrogenase) ELISA Kit 人山梨醇脱氢酶Multi-class antibodies规格： 48T

Anti-Nephrin 去氧抗体Multi-class antibodies规格： 0.1ml

Rhesus antibody Rh IL-27R/TCCR 白细胞介素27受体抗体 规格 0.2ml

SDA(Human salivary duct autoantibody) ELISA Kit 人抗唾液腺导管组织抗体 96T

phospho-P53(Ser6) 英文名称： 磷酸化抑制基因P53抗体 0.1ml

Carboxypeptidase A2 英文名称： 羧肽酶A2抗体 0.1ml

Anti-Nephrin 去氧抗体Multi-class antibodies规格： 0.1ml

CL-0180P388D1(小鼠淋巴样瘤细胞)5×106cells/瓶×2

CD58 Others Human 人 LFA-3 / CD58 人细胞裂解液 (阳性对照)

人间充质干细胞-脂肪裂解物HMSC-ad L

ER-30细胞，人癌细胞 大鼠腺癌细胞系,MADB106细胞 淋巴成纤维细胞Many types of cells包装：5 × 105次方(1ml)

B16-F1(小鼠细胞) 5×106cells/瓶×2

NHEM.f M2 Pellet 正常人表皮黑素细胞团块(少年者)，培养在M2培养液中 > 1 mio.cells 人气管上皮细胞裂解物HTEpiCL

CL-0178OVCAR-3(人细胞)5×106cells/瓶×2

FZD5 Others Mouse 小鼠 Frizzled-5 / FZD5 CHO细胞裂解液 (阳性对照)

人前脂肪细胞-皮下裂解物HPA-s L

小耳猪肺细胞;SEP-L1 癌细胞，MDA-MB-435S细胞 MA891细胞，小鼠癌细胞

人EB病毒转化的B细胞;CGM1

HA Others H2N2 甲型 H2N2 (A/Canada/720/2005) 血凝素HA1 (Hemagglutinin) 人细胞裂解液 (阳性对照)

孕激素受体抗体说明书 STK24 Others Human 人 STK24 / MST3 杆状病毒-昆虫细胞裂解液 (阳性对照)

人肾皮质近曲小管上皮细胞;HK-2

CM-H013人小气道平滑肌细胞完全培养基100mL

MOLT-4细胞，急性淋巴母细胞细胞 髓性细胞,TT细胞 EB病毒转化的人B淋巴细胞(傣族);KM9403

615小鼠T细胞性瘤株;L7712

CD3E Others Human 人 CD3E 人细胞裂解液 (阳性对照)