產(chǎn)品訂購信息：
英文名稱  Anti-Phospho-Rb/P105 RB (Thr821) 

中文名稱   磷酸化視網(wǎng)膜母細(xì)胞瘤相關(guān)蛋白1抗體說明書 

別    名  OSRC; P105 RB; P105RB; PP105; PP110; pRb; RB 1; RB1; RB1 protein; Retinoblastoma 1 (including osteosarcoma); Retinoblastoma 1; Retinoblastoma associated protein; Retinoblastoma related osteosarcoma; Retinoblastoma susceptibility gene; Including osteosarcoma; RB_HUMAN.

濃    度   1mg/1ml

規(guī) 格  0.1ml/100μg

抗體來源   Rabbit

克隆類型   polyclonal

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

產(chǎn)品類型   一抗 磷酸化抗體  

研究領(lǐng)域     細(xì)胞生物 免疫學(xué) 信號(hào)轉(zhuǎn)導(dǎo) 轉(zhuǎn)錄調(diào)節(jié)因子

蛋白分子量  predicted molecular weight: 102kDa

性    狀   Lyophilized or Liquid

免 疫 原 KLH conjugated Synthesised phosphopeptide derived from human Retinoblastoma associated protein around the phosphorylation site of Thr821 

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

磷酸化視網(wǎng)膜母細(xì)胞瘤相關(guān)蛋白1抗體說明書 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 Rb is a tumor suppressor gene which functions as a negative regulator of the cell cycle by interacting with transcription factors including E2F1, PU1, ATF2, UBF, Elf1 and cAbl. This ability of Rb to alter transcription is regulated by phosphorylation catalyzed by the cyclin dependent protein kinases (cdks). Rb is phosphorylated on serine and threonine, but not on tyrosine residues. It forms a complex with SV40 large T antigen, adenovirus E1A, and human papilloma virus 16E. Rb protein may act by regulating transcription and loss of its function leads to uncontrolled cell growth. Aberrations in the Rb gene have been implicated in cancers of breast, colon, prostate, kidney, nasopharynx, and leukemia.

Function : Key regulator of entry into cell division that acts as a tumor suppressor. Promotes G0-G1 transition when phosphorylated by CDK3/cyclin-C. Acts as a transcription repressor of E2F1 target genes. The underphosphorylated, active form of RB1 interacts with E2F1 and represses its transcription activity, leading to cell cycle arrest. Directly involved in heterochromatin formation by maintaining overall chromatin structure and, in particular, that of constitutive heterochromatin by stabilizing histone methylation. Recruits and targets histone methyltransferases SUV39H1, SUV420H1 and SUV420H2, leading to epigenetic transcriptional repression. Controls histone H4 'Lys-20' trimethylation. Inhibits the intrinsic kinase activity of TAF1. Mediates transcriptional repression by SMARCA4/BRG1 by recruiting a histone deacetylase (HDAC) complex to the c-FOS promoter. In resting neurons, transcription of the c-FOS promoter is inhibited by BRG1-dependent recruitment of a phospho-RB1-HDAC1 repressor complex. Upon calcium influx, RB1 is dephosphorylated by calcineurin, which leads to release of the repressor complex (By similarity). In case of viral infections, interactions with SV40 large T antigen, HPV E7 protein or adenovirus E1A protein induce the disassembly of RB1-E2F1 complex thereby disrupting RB1's activity.

Subunit : Interacts with ATAD5. Interacts with PRMT2, CDK1 and CDK2. The hypophosphorylated form interacts with and sequesters the E2F1 transcription factor. Interacts with heterodimeric E2F/DP transcription factor complexes containing TFDP1 and either E2F1, E2F3, E2F4 or E2F5, or TFDP2 and E2F4. The unphosphorylated form interacts with EID1, ARID3B, KDM5A, SUV39H1, MJD2A/JHDM3A and THOC1. Interacts with the N-terminal domain of TAF1. Interacts with AATF, DNMT1, LIN9, LMNA, SUV420H1, SUV420H2, PELP1 and TMPO-alpha. May interact with NDC80. Interacts with GRIP1 and UBR4. Interacts with ARID4A and KDM5B. Interacts with E4F1 and LIMD1. Interacts with SMARCA4/BRG1 AND HDAC1. Interacts with adenovirus E1A protein, HPV E7 protein and SV40 large T antigen. Interacts with PSMA3 and USP4. Interacts (when methylated at Lys-860) with L3MBTL1.

Subcellular Location : Nucleus.

Tissue Specificity : Expressed in the retina.

Post-translational modifications : Phosphorylated by CDK6 and CDK4, and subsequently by CDK2 at Ser-567 in G1, thereby releasing E2F1 which is then able to activate cell growth. Dephosphorylated at the late M phase. SV40 large T antigen, HPV E7 and adenovirus E1A bind to the underphosphorylated, active form of pRb. Phosphorylation at Thr-821 and Thr-826 promotes interaction between the C-terminal domain C and the Pocket domain, and thereby inhibits interactions with heterodimeric E2F/DP transcription factor complexes. Dephosphorylated at Ser-795 by calcineruin upon calcium stimulation. CDK3/cyclin-C-mediated phosphorylation at Ser-807 and Ser-811 is required for G0-G1 transition. Phosphorylated by CDK1 and CDK2 upon TGFB1-mediated apoptosis.

N-terminus is methylated by METTL11A/NTM1. Monomethylated at Lys-860 by SMYD2, promoting interaction with L3MBTL1.

DISEASE : Defects in RB1 are the cause of childhood cancer retinoblastoma (RB) [MIM:180200]. RB is a congenital malignant tumor that arises from the nuclear layers of the retina. It occurs in about 1:20'000 live births and represents about 2% of childhood malignancies. It is bilateral in about 30% of cases. Although most RB appear sporadically, about 20% are transmitted as an autosomal dominant trait with incomplete penetrance. The diagnosis is usually made before the age of 2 years when strabismus or a gray to yellow reflex from pupil ('cat eye') is investigated.

Defects in RB1 are a cause of susceptibility to bladder cancer (BLC) [MIM:109800]. A malignancy originating in tissues of the urinary bladder. It often presents with multiple tumors appearing at different times and at different sites in the bladder. Most bladder cancers are transitional cell carcinomas. They begin in cells that normally make up the inner lining of the bladder. Other types of bladder cancer include squamous cell carcinoma (cancer that begins in thin, flat cells) and adenocarcinoma (cancer that begins in cells that make and release mucus and other fluids). Bladder cancer is a complex disorder with both genetic and environmental influences.

Defects in RB1 are a cause of osteogenic sarcoma (OSRC) [MIM:259500].

Similarity : Belongs to the retinoblastoma protein (RB) family.

Database links : UniProtKB/Swiss-Prot: P06400.2

MIP2 (myocardial ischemic preconditioning upregulated protein 2) 巨噬細(xì)胞蛋白2抗原Multi-class antibodies規(guī)格： 0.5mg

Anti-Angiopoietin 1 生成素-1抗體Multi-class antibodies規(guī)格： 0.1ml

Rhesus antibody Rh phospho-Beta-catenin(Tyr86) 磷酸化β 連環(huán)素蛋白抗體 規(guī)格 0.1ml

包被緩沖液(PH9.6) 100ml 進(jìn)口分裝

VTG 英文名稱： 魚、青鳉魚卵黃蛋白原抗體 0.2ml

DLGAP4 英文名稱： PSD95結(jié)合蛋白4抗體 0.2ml

Anti-Angiopoietin 1 生成素-1抗體Multi-class antibodies規(guī)格： 0.1ml

Anti-phospho-P53(pSer392)/FITC 熒光素標(biāo)記抗磷酸化抑制基因P53抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rabbit Anti-mouse IgG/Gold 金標(biāo)記兔抗小鼠IgG(10nm/15nm)Multi-class antibodies規(guī)格： 0.5ml

纖維連結(jié)蛋白抗體 Anti-FN 0.1ml

Goat Anti-Mouse IgG/Alexa Fluor 350 Alexa Fluor 350標(biāo)記的羊抗小鼠IgG 0.1ml

FNDC4 英文名稱： Ⅲ型纖維連接蛋白域蛋白4抗體 0.2ml

Rhesus antibody Rh phospho-STAT5a(Tyr694) 磷酸化信號(hào)轉(zhuǎn)導(dǎo)和轉(zhuǎn)錄激活因子5a抗體 規(guī)格 0.1ml

Rabbit Anti-mouse IgG/Gold 金標(biāo)記兔抗小鼠IgG(10nm/15nm)Multi-class antibodies規(guī)格： 0.5ml

Syntenin 1 英文名稱： 多配體聚糖結(jié)合蛋白1抗體 0.2ml

Endostatin 英文名稱： 內(nèi)皮抑素/內(nèi)皮他丁抗體 0.1ml

組織蛋白酶G抗體 Anti-CTSG/CG 0.1ml

Anti-UCP-3/FITC 熒光素標(biāo)記線粒體脫偶連蛋白3抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh Phospho-Histone H3(Ser28) 磷酸化組蛋白H3抗體 規(guī)格 0.1ml

CXCR6(CXC-chemokine receptor 6) peptide 細(xì)胞表面趨化因子受體6抗原Multi-class antibodies規(guī)格： 0.5mg

RDFs, 大鼠真皮成纖維細(xì)胞

16HBE 人上皮細(xì)胞

MIF Others Mouse 小鼠 MIF / Migration Inhibitory Factor 人細(xì)胞裂解液 (陽性對(duì)照)

CL-0263BGC-803(人細(xì)胞)5×106cells/瓶×2

人肝內(nèi)膽管上皮細(xì)胞cDNAHIBEpiC cDNA

Asp2人胚胎腎細(xì)胞轉(zhuǎn)化細(xì)胞;FC33 人腎小球內(nèi)皮細(xì)胞完全培養(yǎng)基 100mL

CL-0148MADB106(大鼠癌細(xì)胞)5×106cells/瓶×2

SELL Others Cynomolgus 食蟹猴 CD62L / L-Selectin / SELL 人細(xì)胞裂解液 (陽性對(duì)照)

人上皮細(xì)胞裂解物HPEpiCL

1590細(xì)胞，人癌細(xì)胞系 小鼠,EL-4細(xì)胞 腦平滑肌細(xì)胞Many types of cells包裝：5 × 105次方(1ml)

Beta-TC-6(小鼠胰島素瘤胰島β細(xì)胞) 5×106cells/瓶×2

HCASMC Pellet 人平滑肌細(xì)胞團(tuán)塊 > 1 mio.cells 人成纖維細(xì)胞-心室裂解物HCF-av L

磷酸化視網(wǎng)膜母細(xì)胞瘤相關(guān)蛋白1抗體說明書 RDFs, 大鼠真皮成纖維細(xì)胞

16HBE 人上皮細(xì)胞

MIF Others Mouse 小鼠 MIF / Migration Inhibitory Factor 人細(xì)胞裂解液 (陽性對(duì)照)

CL-0263BGC-803(人細(xì)胞)5×106cells/瓶×2

人肝內(nèi)膽管上皮細(xì)胞cDNAHIBEpiC cDNA

Asp2人胚胎腎細(xì)胞轉(zhuǎn)化細(xì)胞;FC33 人腎小球內(nèi)皮細(xì)胞完全培養(yǎng)基 100mL

抗體的生物素化標(biāo)記實(shí)驗(yàn)要點(diǎn)：

1. 磷酸化視網(wǎng)膜母細(xì)胞瘤相關(guān)蛋白1抗體說明書 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。

抗體的鑒定：

1）磷酸化視網(wǎng)膜母細(xì)胞瘤相關(guān)蛋白1抗體說明書 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?？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。