抗體的生物素化標記實驗要點：

1. 環(huán)指蛋白123抗體規(guī)格 如在反應混合液中有疊氮鈉或游離氨基存在,會抑制標記反應。因此,蛋白質(zhì)在反應前要對 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,選擇不當則影響標記的效率,應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)反應后,應充分透析,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,故對某些細胞可導致高本底。

產(chǎn)品訂購信息：
英文名稱  Anti-RNF123 

中文名稱   環(huán)指蛋白123抗體規(guī)格 

別    名  E3 ubiquitin protein ligase RNF123; Kip1 ubiquitination promoting complex protein 1; KPC 1; KPC1; Ring finger protein 123; RNF 123; Ubiquitin ligase;

濃    度   1mg/1ml

規(guī) 格  0.2ml/200μg

抗體來源   Rabbit

克隆類型   polyclonal

交叉反應   Human, Mouse, Rat, Dog, Pig 

產(chǎn)品類型   一抗  

研究領域    細胞生物 免疫學

蛋白分子量  predicted molecular weight: 149kDa

性    狀   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human RNF123 

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

環(huán)指蛋白123抗體規(guī)格 產(chǎn)品應用    WB=1:100-500 ELISA=1:500-1000 IHC-P=1:100-500 IHC-F=1:100-500 IF=1:50-200

（石蠟切片需做抗原修復） 

 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 The RING-type zinc finger motif is present in a number of viral and eukaryotic proteins and is made of a conserved cysteine-rich domain that is able to bind two zinc atoms. Proteins that contain this conserved domain are generally involved in protein-protein interactions and protein-DNA interactions. RNF123 (RING finger protein 123), also known as KPC1 (Kip1 (p27) ubiquitination-promoting complex protein 1) or FP1477, contains one RING-type zinc finger domain and one SPRY domain. Localizing to the cytoplasm, RNF123 functions as the catalytic component of the KPC complex that acts as an E3 ubiquitin-protein ligase. Specifically, RNF123 is essential for the ubiquitination and subsequent degradation of p27 during the cell cycle G1 phase. Via its N-terminus, RNF123 is known to interact with GBDR1 (another component of the KPC) and p27 (a cyclin-dependent kinase inhibitor). Due to alternative splicing events, two isoforms exist for RNF123.

Function : Catalytic subunit of the KPC complex that acts as E3 ubiquitin-protein ligase. Required for poly-ubiquitination and proteasome-mediated degradation of CDKN1B during G1 phase of the cell cycle.

Subunit : Component of the KPC complex composed of RNF123/KPC1 and UBAC1/KPC2. Interacts with UBAC1 and CDKN1B via its N-terminal domain.

Subcellular Location : Cytoplasmic

Post-translational modifications : Ubiquitinated, leading to its degradation. Deubiquitinated by USP19, thereby stimulating CDKN1B ubiquitin-dependent degradation (By similarity).

Similarity : Contains 1 B30.2/SPRY domain.

Contains 1 RING-type zinc finger.

Database links : UniProtKB/Swiss-Prot: Q5XPI4.1

抗體的鑒定：

1）環(huán)指蛋白123抗體規(guī)格 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,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抗原或近似抗原物質(zhì)的識別能力?？贵w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率來表示。交叉反應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。

FGF4/HSTF1/HBGF-4(Fibroblast Growth Factor4) 纖維母細胞生長因子4抗原Multi-class antibodies規(guī)格： 0.5mg

腦素/利肽抗體 Anti-BNP 0.1ml

Rhesus antibody Rh phospho-E2F1 (Ser332) 磷酸化轉(zhuǎn)錄因子E2F-1抗體 規(guī)格 0.1ml

纖維素膜(30cm×3m 0.45um) 1卷 Whatman

ZNF281 英文名稱： 鋅指蛋白281抗體 0.2ml

DEF6/IBP 英文名稱： 干擾素調(diào)節(jié)因子4結(jié)合蛋白抗體 0.2ml

腦素/利肽抗體 Anti-BNP 0.1ml

Anti-phospho-SHC (Tyr239/240)/FITC 熒光素標記磷酸化SH2結(jié)構(gòu)域轉(zhuǎn)化蛋白1抗體IgGMulti-class antibodies規(guī)格： 0.2ml

VTG(vitellogenin) 青鳉魚卵黃蛋白Multi-class antibodies規(guī)格： 0.1mg

細胞核因子/k基因結(jié)合核因子抗體 Anti-NFKBp65 0.1ml

SCUBE1 英文名稱： 新型血小板相關內(nèi)皮分泌蛋白SCUBE1抗體 0.2ml

EV71 polyprotein VP1 英文名稱： 腸道病毒71型抗體 0.2ml

Rhesus antibody Rh phospho-p95 NBS1(Ser343) 磷酸化滑膜細胞凋亡抑制物1抗體 規(guī)格 0.1ml

VTG(vitellogenin) 青鳉魚卵黃蛋白Multi-class antibodies規(guī)格： 0.1mg

ZBTB7C 英文名稱： 鋅指蛋白ZBTB7C抗體 0.2ml

DNTNP 英文名稱： 背神經(jīng)管核蛋白抗體 0.2ml

膜粘連蛋白 Ⅱ抗體 Anti-Annexin Ⅱ 0.1ml

Anti-ZNF474/FITC 熒光素標記鋅指蛋白474抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh phospho-ERK5(Ser496) 磷酸化細胞外信號調(diào)節(jié)激酶5抗體 規(guī)格 0.1ml

DRD5 receptor(dopamine D5 receptor) 多巴胺受體-D5抗原Multi-class antibodies規(guī)格： 0.5mg

TNFSF11 Protein Human 重組人 RANKL / OPGL / TNFSF11 / CD254 蛋白 (Fc 標簽)

HEC-1-A 人子宮內(nèi)膜腺癌細胞

LSAMP Others Human 人 LSAMP 人細胞裂解液 (陽性對照)

Caki-1人腎透明細胞癌細胞 Caki-1 in human renal clear cell carcinoma of skin metastasis cell McCOY's *+10%FBS

人胚腎細胞;293 [HEK-293]

大鼠神經(jīng)黑質(zhì)細胞(RNsn)(1×106) BxPC-3, 人原位*腺癌細胞株 Human

CL-0398NCI-H2452(人間皮瘤細胞)5×106cells/瓶×2

CD22 Others Mouse 小鼠 Siglec-2 / CD22 人細胞裂解液 (陽性對照)

人腦膜細胞RNAHMC miRNA5 μg

家兔腎細胞;RABK3 細胞，COLO 205細胞 U14-GFP細胞，綠色熒光蛋白標記小鼠子細胞

人肺轉(zhuǎn)化細胞;AE1201

HA Others H1N1 甲型 H1N1 (A/Solomon Islands/3/2006) 血凝素 (Hemagglutinin / HA) 人細胞裂解液 (陽性對照)

環(huán)指蛋白123抗體規(guī)格 TNFSF11 Protein Human 重組人 RANKL / OPGL / TNFSF11 / CD254 蛋白 (Fc 標簽)

HEC-1-A 人子宮內(nèi)膜腺癌細胞

LSAMP Others Human 人 LSAMP 人細胞裂解液 (陽性對照)

Caki-1人腎透明細胞癌細胞 Caki-1 in human renal clear cell carcinoma of skin metastasis cell McCOY's *+10%FBS

人胚腎細胞;293 [HEK-293]

大鼠神經(jīng)黑質(zhì)細胞(RNsn)(1×106) BxPC-3, 人原位*腺癌細胞株 Human