抗體的生物素化標(biāo)記實驗要點：

1. 凋亡相關(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,會抑制標(biāo)記反應(yīng)。因此,蛋白質(zhì)在反應(yīng)前要對 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-氨基的密度會有所不同,選擇不當(dāng)則影響標(biāo)記的效率,應(yīng)先用幾個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)合位點時,或位于酶的催化位點時,生物素化會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,如羰基、氨基、巰基、異咪唑基及苯酚基,也可與糖基共價結(jié)合；

7.交聯(lián)反應(yīng)后,應(yīng)充分透析,否則,殘余的生物素會對生物素化抗體與親和素的結(jié)合產(chǎn)生競爭作用；

8.在細胞的熒光標(biāo)記實驗中,中和親和素的本底低,但由于鏈霉親和素含有少量正電荷,故對某些細胞可導(dǎo)致高本底。

產(chǎn)品訂購信息：
英文名稱  Anti-RYBP/APAP1 

中文名稱   凋亡相關(guān)蛋白1抗體說明書 

別    名  AAP 1; AAP1; APAP 1; APAP-1; APAP1; Apoptin associating protein 1; Apoptin-associating protein 1; cb337; Death Effector Domain Associated Factor; Death effector domain-associated factor; Death effector domain-associated factor B; DED associated factor; DED-associated factor; DED-associated factor B; DEDAF; RING1 and YY1 binding protein a; RING1 and YY1 binding protein; RING1 and YY1-binding protein; RING1 and YY1-binding protein B; Ring1 interactor RYBP; RYBP; RYBP_HUMAN; rybpa; YEAF 1; YEAF1; YY1 and E4TF1 associated factor 1; YY1 and E4TF1-associated factor 1; YY1- and E4TF1/GABP-associated factor 1.

濃    度   1mg/1ml

規(guī) 格  0.2ml/200μg

抗體來源   Rabbit

克隆類型   polyclonal

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

產(chǎn)品類型   一抗  

研究領(lǐng)域    細胞生物 信號轉(zhuǎn)導(dǎo) 細胞凋亡

蛋白分子量  predicted molecular weight: 24kDa

性    狀   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human AAP1 

亞    型   IgG

純化方法   affinity purified by Protein A

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

凋亡相關(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 May be implicated in the regulation of the transcription as a repressor of the transcriptional activity of E4TF1. In tumor cell lines, may induce apoptosis.

Function : nhibits ubiquitination and subsequent degradation of TP53, and thereby plays a role in regulating transcription of TP53 target genes. May be implicated in the regulation of the transcription as a repressor of the transcriptional activity of E4TF1. May bind to DNA. Promotes apoptosis.

Subunit : Monomer. Interacts with apoptin, DEDD, FADD, CASP8, CASP10, YY1 and GABPB1. Component of repressive BCOR complex containing Polycomb group subcomplex at least composed of BCOR, PCGF1, RING1 and RNF2/RING2. Together with GABPB1 and YY1, it forms a ternary complex, probably being the bridge factor between these two transcription factors. Interacts with CBX2, YAF2, RING1 and RNF2. Interacts with ubiquitin and ubiquitinated proteins. Interacts with ubiquitinated histone H2A (By similarity). Interacts with MDM2, and thereby inhibits ubiquitination of TP53. Identified in a ternary complex containing MDM2, TP53 and RYBP.

Subcellular Location : Nucleus. Cytoplasm. Nucleus, nucleoplasm (By similarity). Note=Primarily found in the nucleus. Detected in a punctate pattern likely to represent Polycomb group (PcG) bodies

Tissue Specificity : Widely expressed with highest levels in lymphoid tissues and placenta.

Post-translational modifications : Monoubiquitinated

Similarity : Contains 1 RanBP2-type zinc finger.

Database links : UniProtKB/Swiss-Prot: Q8N488.2

抗體的鑒定：

1）凋亡相關(guān)蛋白1抗體說明書 抗體的效價鑒定：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。不同的抗原制備的抗體,要求的效價不一。鑒定效價的方法很多,包括有試管凝集反應(yīng),瓊脂擴散試驗,酶聯(lián)免疫吸附試驗等。常用的抗原所制備的抗體一般都有約成的鑒定效價的方法,以資比較。如制備抗抗體的效價,一般就采用瓊脂擴散試驗來鑒定。

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)的識別能力。抗體的特異性高,它的識別能力就強。衡量特異性通常以交叉反應(yīng)率來表示。交叉反應(yīng)率可用競爭抑制試驗測定。以不同濃度抗原和近似抗原分別做競爭抑制曲線,計算各自的結(jié)合率,求出各自在IC50時的濃度,并按公式計算交叉反應(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,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親和力的測定對抗體的篩選,確定抗體的用途,驗證抗體的均一性等均有重要意義。

Histone H1b(Histone H1.4) 組蛋白H1b抗原Multi-class antibodies規(guī)格： 0.5mg

Anti-5-HTR2A 5-羥色胺受體2A抗體Multi-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh Phospho-c-Kit(Tyr721) 磷酸化原癌基因c-kit抗體 規(guī)格 0.1ml

羊抗雞IgG血清 0.5ml 國產(chǎn)

XIAP 英文名稱： X-連鎖凋亡蛋白/性連鎖凋亡抑制蛋白抗體 0.1ml

DNA PKcs 英文名稱： DNA依賴蛋白激酶催化亞基抗體 0.2ml

Anti-5-HTR2A 5-羥色胺受體2A抗體Multi-class antibodies規(guī)格： 0.2ml

Anti-TP II A/TOP2a/FITC 熒光素標(biāo)記DNA拓普西異構(gòu)酶ⅡA抗體IgGMulti-class antibodies規(guī)格： 0.2ml

SAP-1 (Synapse associated protein-I) peptide 突觸相關(guān)蛋白-1(多肽抗原)Multi-class antibodies規(guī)格： 0.5mg

酪氨酸激酶A抗體 Anti-TrK A 0.1ml

SLU7 英文名稱： SLU7蛋白抗體 0.2ml

EN2 英文名稱： 同源盒蛋白轉(zhuǎn)錄因子EN2抗體 0.1ml

Rhesus antibody Rh phospho-IRS1(Ser612) 磷酸化受體底物-1抗體 規(guī)格 0.1ml

SAP-1 (Synapse associated protein-I) peptide 突觸相關(guān)蛋白-1(多肽抗原)Multi-class antibodies規(guī)格： 0.5mg

SARP1 英文名稱： 分泌細胞凋亡相關(guān)蛋白1抗體 0.2ml

ESRP2 英文名稱： 上皮粘連調(diào)控蛋白ESRP2抗體 0.2ml

轉(zhuǎn)鐵蛋白受體抗體 Anti-CD71/TFR 0.1ml

Anti-HSP-70/RBITC 羅丹明標(biāo)記熱休克蛋白-70抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh phospho-GATA1(ser142) 磷酸化珠蛋白轉(zhuǎn)錄因子1抗體 規(guī)格 0.1ml

CG6856-PA peptide CG6856-PA抗原Multi-class antibodies規(guī)格： 0.5mg

EPO Protein Rat 重組大鼠 Erythropoietin / EPO 蛋白 (His 標(biāo)簽)

MG-63 人成細胞

CD93 Others Human 人 CD93 / C1QR1 人細胞裂解液 (陽性對照)

SK-MES-1人肺鱗癌細胞 SK-MES-1 human lung squamous carcinoma cells MEM培養(yǎng)基(GIBCO)+10%FBS

人導(dǎo)管瘤細胞;UACC812

SV40T轉(zhuǎn)化的人胚腎細胞(亞系);293T/17 人上皮細胞完全培養(yǎng)基 100mL

CL-0267CNE-2Z(人細胞)5×106cells/瓶×2

PPT1 Others Human 人 PPT1 / Palmitoyl-protein thioesterase 1 人細胞裂解液 (陽性對照)

人主動脈內(nèi)皮細胞cDNAHAEC cDNA

豬腎傳代細胞;IBRS-2 人B淋巴母細胞，HMy2.CIR細胞 LⅡ細胞，KM小鼠網(wǎng)織細胞瘤株

穩(wěn)定表達EBNA1的人胚腎細胞;293E

HA Others H5N1 甲型 H5N1 (A/Egypt/2321-NAMRU3/2007) 血凝素HA1 (Hemagglutinin) 人細胞裂解液 (陽性對照)

凋亡相關(guān)蛋白1抗體說明書 EPO Protein Rat 重組大鼠 Erythropoietin / EPO 蛋白 (His 標(biāo)簽)

MG-63 人成細胞

CD93 Others Human 人 CD93 / C1QR1 人細胞裂解液 (陽性對照)

SK-MES-1人肺鱗癌細胞 SK-MES-1 human lung squamous carcinoma cells MEM培養(yǎng)基(GIBCO)+10%FBS

人導(dǎo)管瘤細胞;UACC812

SV40T轉(zhuǎn)化的人胚腎細胞(亞系);293T/17 人上皮細胞完全培養(yǎng)基 100mL