抗體的生物素化標(biāo)記實(shí)驗(yàn)要點(diǎn)：

1. 磷酸化孕激素受體抗體規(guī)格 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(lài)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。

產(chǎn)品訂購(gòu)信息：
英文名稱(chēng)  Anti-Phospho-Progesterone Receptor(Ser190) 

中文名稱(chēng)   磷酸化孕激素受體抗體規(guī)格 

別    名  PGR(phospho S190); NR3C3; Nuclear receptor subfamily 3 group C member 3; PGR; PR; PRA; PRB; Progesterone receptor; Progestin receptor form A; Progestin receptor form B; PRGR_RAT.

濃    度   1mg/1ml

規(guī) 格  0.1ml/100μg

抗體來(lái)源   Rabbit

克隆類(lèi)型   polyclonal

交叉反應(yīng)   Mouse, Rat, Cow 

產(chǎn)品類(lèi)型   一抗 磷酸化抗體  

研究領(lǐng)域     染色質(zhì)和核信號(hào) 信號(hào)轉(zhuǎn)導(dǎo) 激酶和磷酸酶

蛋白分子量  predicted molecular weight: 99kDa

性    狀   Lyophilized or Liquid

免 疫 原 KLH conjugated Synthesised phosphopeptide derived from rat Progesterone Receptor around the phosphorylation site of Ser190 

亞    型   IgG

純化方法   affinity purified by Protein A

儲(chǔ) 存 液   0.01M PBS, pH 7.4 with 10 mg/ml BSA and 0.1% Sodium azide

磷酸化孕激素受體抗體規(guī)格 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=1:100-500

（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 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 UNC4*. 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.

Isoform A: Nucleus. Cytoplasm. Note=Mainly nuclear.

Tissue Specificity : Isoform A: Nucleus. Cytoplasm. Note=Mainly nuclear.

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.

Ubiquitination is hormone-dependent and represses sumoylation on the same site. Promoted by MAPK-mediated phosphorylation on Ser-294.

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

Contains 1 nuclear receptor DNA-binding domain.

Database links : UniProtKB/Swiss-Prot: Q63449.1

類(lèi)固醇受體（Steroid Receptors）孕激素受體是一類(lèi)位于孕酮靶組織細(xì)胞內(nèi)或細(xì)胞表面的特異蛋白質(zhì)，特異地與孕酮結(jié)合，所形成的細(xì)胞溶質(zhì)孕酮-受體復(fù)合物隨后與細(xì)胞核內(nèi)的DNA結(jié)合，以啟動(dòng)蛋白質(zhì)生物合成。孕酮受體有A和B兩種，受雌激素誘導(dǎo)，半壽期很短。

抗體的鑒定：

1）磷酸化孕激素受體抗體規(guī)格 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。

Anti-CD26/ADCP2/DPP IV /FITC 熒光素標(biāo)記CD26抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Anti-H-ras/FITC 熒光素標(biāo)記原癌基因H-ras抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh CDC123/C10orf7 細(xì)胞分裂周期蛋白CDC123抗體 規(guī)格 0.2ml

Polycystin 2(polycystic kidney disease 2) 蛋白2抗體 0.5mg

IFITM2 英文名稱(chēng)： 干擾素誘導(dǎo)跨膜蛋白2抗體 0.2ml

Rhesus antibody Rh TCF7L2 轉(zhuǎn)錄因子7類(lèi)似物2抗體 規(guī)格 0.1ml

Anti-H-ras/FITC 熒光素標(biāo)記原癌基因H-ras抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Anti-KLK14/FITC 熒光素標(biāo)記激肽釋放酶14抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Anti-phosphor-SMAD(p-Ser425)、phosphor-Mothers against decapentaplegic/FITC 熒光素標(biāo)記磷酸化SMAD抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh ALG11 天門(mén)冬酰胺連接糖基化11抗體 規(guī)格 0.2ml

Rabbit Anti-human IgG/PE-Cy3 PE-Cy3標(biāo)記的兔抗人IgG 0.1ml

GRB 14 英文名稱(chēng)： 生長(zhǎng)因子受體結(jié)合蛋白14抗體 0.1ml

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Anti-phosphor-SMAD(p-Ser425)、phosphor-Mothers against decapentaplegic/FITC 熒光素標(biāo)記磷酸化SMAD抗體IgGMulti-class antibodies規(guī)格： 0.2ml

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Anti-Urocortin 新型活性因子UCN抗體Multi-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh G-CSFR/CD114 粒細(xì)胞-巨噬細(xì)胞集落刺激因子3受體抗體 規(guī)格 0.1ml

Casp-9 ELISA Kit 大鼠胱天蛋白酶9 96T

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APH1a 英文名稱(chēng)： 早老素γ分泌酶抗體 0.2ml

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CL-0080WB-F344(大鼠肝上皮樣干細(xì)胞)5×106cells/瓶×2

AKT3 Others Human 人 AKT3 桿狀病毒-昆蟲(chóng)細(xì)胞裂解液 (陽(yáng)性對(duì)照)

SMCFectagen? 平滑肌細(xì)胞轉(zhuǎn)染試劑盒

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家兔腎細(xì)胞;RABK3

SFRP4 Others Mouse 小鼠 sFRP4 人細(xì)胞裂解液 (陽(yáng)性對(duì)照)

CL-0079ES-2(人卵巢透明細(xì)胞癌細(xì)胞)5×106cells/瓶×2

IL17RC Others Human 人 IL17RC 人細(xì)胞裂解液 (陽(yáng)性對(duì)照)

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CF Protein Human 重組人 CF 蛋白 (His 標(biāo)簽)

SF126(人細(xì)胞) 5×106cells/瓶×2 RD(惡性胚胎橫紋肌瘤)

磷酸化孕激素受體抗體規(guī)格 人脈絡(luò)絲成纖維細(xì)胞 (HCPF)( 5×105 )

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腸平滑肌細(xì)胞Many types of cells包裝：5 × 105次方(1ml)

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DH82 狗腎惡性癥細(xì)胞

CD19 Others Human 人 CD19 / Leu-12 人細(xì)胞裂解液 (陽(yáng)性對(duì)照)