抗體的生物素化標(biāo)記實(shí)驗(yàn)要點(diǎn)：

1. 表觀抑制因子Polycomb家族成員PCGFl蛋白抗體費(fèi)用 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)不同的分子比來篩選最適條件；

3.用NHSB量過量也是不利的,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)品訂購信息：
英文名稱  Anti-PCGF1 

中文名稱   表觀抑制因子Polycomb家族成員PCGFl蛋白抗體費(fèi)用 

別    名  PCGF1_HUMAN; PCGF1; Nervous system Polycomb-1; NSPc1; Polycomb group RING finger protein 1; RING finger protein 68; RNF68.

濃    度   1mg/1ml

規(guī) 格  0.2ml/200μg 1ml/1mg

抗體來源   Rabbit

克隆類型   polyclonal

交叉反應(yīng)   Human, Mouse, Rat, Dog, Cow, Horse, Rabbit 

產(chǎn)品類型   一抗  

研究領(lǐng)域    細(xì)胞生物 細(xì)胞周期蛋白 細(xì)胞分化

蛋白分子量  predicted molecular weight: 30kDa

性    狀   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from hu PCGF1 

亞    型   IgG

純化方法   affinity purified by Protein A

儲(chǔ) 存 液   Preservative: 15mM Sodium Azide, Constituents: 1% BSA, 0.01M PBS, pH 7.4

表觀抑制因子Polycomb家族成員PCGFl蛋白抗體費(fèi)用 產(chǎn)品應(yīng)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 PCGF1 is a component of the Polycomb group (PcG) multiprotein BCOR complex, a complex required to maintain the transcriptionally repressive state of some genes, such as BCL6 and the cyclin-dependent kinase inhibitor, CDKN1A. It represses CDKN1A expression by binding to its promoter, and this repression is dependent on the retinoic acid response element (RARE element). PCGF1 als promotes cell cycle progression and also enhances cell proliferation, thus it may have a positive role in tumor cell growth.

Function : Component of the Polycomb group (PcG) multiprotein BCOR complex, a complex required to maintain the transcriptionally repressive state of some genes, such as BCL6 and the cyclin-dependent kinase inhibitor, CDKN1A. Transcriptional repressor that may be targeted to the DNA by BCL6; this transcription repressor activity may be related to PKC signaling pathway. Represses CDKN1A expression by binding to its promoter, and this repression is dependent on the retinoic acid response element (RARE element). Promotes cell cycle progression and enhances cell proliferation as well. May have a positive role in tumor cell growth by down-regulating CDKN1A. Component of a Polycomb group (PcG) multiprotein PRC1-like complex, a complex class required to maintain the transcriptionally repressive state of many genes, including Hox genes, throughout development. PcG PRC1 complex acts via chromatin remodeling and modification of histones; it mediates monoubiquitination of histone H2A 'Lys-119', rendering chromatin heritably changed in its expressibility.

Subunit : Component of repressive BCOR complex containing Polycomb group subcomplex at least composed of RYBP, RING1 and RNF2/RING2. Specifically interacts with BCOR, RING1 and RNF2/RING2. Component of a PRC1-like complex. Interacts with CBX6, CBX7 and CBX8.

Subcellular Location : Nucleus.

Similarity : Contains 1 RING-type zinc finger.

Database links : UniProtKB/Swiss-Prot: Q9BSM1.2

抗體的鑒定：

1）表觀抑制因子Polycomb家族成員PCGFl蛋白抗體費(fèi)用 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。抗體的特異性高,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-CK18(C-term) /FITC 熒光素標(biāo)記細(xì)胞角蛋白18抗體(C端)IgGMulti-class antibodies規(guī)格： 0.2ml

Anti-Phospho-IRAK1 (Thr209) /FITC 熒光素標(biāo)記磷酸化白介素-1受體相關(guān)激酶1抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh CCL27/CTACK 皮膚T細(xì)胞虜獲趨化因子抗體 規(guī)格 0.2ml

IL-23R(Interleukin-23 receptor) 白介素-23受體抗原 0.5mg

HPV18 E7 英文名稱： 人類狀瘤病毒18 E7抗體 0.2ml

Rhesus antibody Rh SPARCL1/Ecm2 細(xì)胞外基質(zhì)蛋白2抗體 規(guī)格 0.2ml

Anti-Phospho-IRAK1 (Thr209) /FITC 熒光素標(biāo)記磷酸化白介素-1受體相關(guān)激酶1抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Anti-MRP4/ABCC4 /FITC 熒光素標(biāo)記多藥耐藥相關(guān)蛋白4抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Anti-Phospho-Tuberin/TSC2 (Tyr1571)/FITC 熒光素標(biāo)記磷酸化馬鈴薯球蛋白(性硬化)抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh 2,4-D(24D1) 2，4-二氯苯氧乙酸(除草劑)單克隆抗體 規(guī)格 0.1ml

Mouse Anti-rat IgG/PE PE標(biāo)記的小鼠抗大鼠IgG 0.1ml

phospho-GluA4 (Ser862) 英文名稱： 磷酸化離子型谷酸受體4抗體 0.1ml

Rhesus antibody Rh PSMD8 蛋白酶調(diào)解因子8抗體 規(guī)格 0.2ml

Anti-Phospho-Tuberin/TSC2 (Tyr1571)/FITC 熒光素標(biāo)記磷酸化馬鈴薯球蛋白(性硬化)抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Ntn1(Human Netrin-1) ELISA Kit 人軸突生長(zhǎng)誘向因子1Multi-class antibodies規(guī)格： 48T

Anti-phospho-PAK1(Ser) 磷酸化p21激活激酶1抗體Multi-class antibodies規(guī)格： 0.1ml

Rhesus antibody Rh HPGD/PGDH1/Prostaglandin dehydrogenase 1 素脫氫酶1抗體 規(guī)格 0.1ml

TA(Human anti-teichoic acid antibody) ELISA Kit 人抗磷壁酸抗體 96T

Phospho-PKR (Thr446) 英文名稱： 磷酸化蛋白激酶R抗體 0.1ml

CD66a 英文名稱： 癌胚抗原相關(guān)細(xì)胞粘附分子1抗體 0.2ml

Anti-phospho-PAK1(Ser) 磷酸化p21激活激酶1抗體Multi-class antibodies規(guī)格： 0.1ml

CL-0381MDA-MB-175VII(人導(dǎo)管癌細(xì)胞)5×106cells/瓶×2

LIPF Others Human 人 Gasic lipase / LIPF 桿狀病毒-昆蟲細(xì)胞裂解液 (陽性對(duì)照)

人心肌細(xì)胞-心室總RNAHCF-av NA

大耳山羊腎細(xì)胞;LDG-2 皮下結(jié)締組織細(xì)胞，A9細(xì)胞 CT26.WT[CT26WT]細(xì)胞，小鼠細(xì)胞

人胚肺二倍體細(xì)胞;KMB-17

HA Others H11N2 甲型 H11N2 (A/duck/Yangzhou/906/2002) 血凝素HA1 (Hemagglutinin) 人細(xì)胞裂解液 (陽性對(duì)照)

CL-0379MCF 7B(人癌細(xì)胞)5×106cells/瓶×2

HDAC8 Others Human 人 HDAC8 / HDACL1 桿狀病毒-昆蟲細(xì)胞裂解液 (陽性對(duì)照)

人心肌細(xì)胞-總RNAHCM-a NA

THP-1細(xì)胞，人單核細(xì)胞 人,H1299細(xì)胞 A549/DDP, 人肺腺癌耐藥細(xì)胞株

家貓腎細(xì)胞;CATK1

HA Others H3N2 甲型 H3N2 (A/Brisbane/10/2007) 血凝素 (Hemagglutinin / HA) 人細(xì)胞裂解液 (陽性對(duì)照)

表觀抑制因子Polycomb家族成員PCGFl蛋白抗體費(fèi)用 GAL2 Others Human 人 GAL2 / GalNAc-T2 人細(xì)胞裂解液 (陽性對(duì)照)

二氫葉酸還原酶缺陷型中國倉鼠卵巢細(xì)胞;CHO/dhFr-

CL-0053Calu-1(人細(xì)胞)5×106cells/瓶×2

ZR-75-30細(xì)胞，人癌細(xì)胞系 人T淋巴細(xì)胞(CD8+),Molt-4細(xì)胞 tTA基因修飾的小鼠*癌細(xì)胞(B類);Pan02-CAG-tTA-4C5

BRL-3A(大鼠正常肝細(xì)胞) 5×106cells/瓶×2

CPE Others Human 人 CPE 人細(xì)胞裂解液 (陽性對(duì)照)