抗體的生物素化標(biāo)記實(shí)驗(yàn)要點(diǎn)：

1. 蓬亂蛋白2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)生競爭作用；

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-shevelled 2 

中文名稱   蓬亂蛋白2抗體規(guī)格 

別    名  Dishevelled 2 (homologous to Drosophila dsh); Dishevelled dsh homolog 2; Dishevelled-2; Dishevelled2; DSH homolog 2; DVL 2; Dvl2; DVL2_HUMAN; Segment polarity protein dishevelled homolog DVL 2; Segment polarity protein dishevelled homolog DVL-2; Segment polarity protein dishevelled homolog DVL2.

濃    度   1mg/1ml

規(guī) 格  0.1ml/100μg 0.2ml/200μg

抗體來源   Rabbit

克隆類型   polyclonal

交叉反應(yīng)   Human, Mouse, Rat 

產(chǎn)品類型   一抗  

研究領(lǐng)域    細(xì)胞生物 免疫學(xué) 信號(hào)轉(zhuǎn)導(dǎo) 干細(xì)胞 細(xì)胞凋亡

蛋白分子量  predicted molecular weight: 81kDa

性    狀   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human shevelled 2 

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

蓬亂蛋白2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 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 This gene encodes a member of the dishevelled (dsh) protein family. The vertebrate dsh proteins have approximately 40% amino acid sequence similarity with Drosophila dsh. This gene encodes a 90-kD protein that undergoes posttranslational phosphorylation to form a 95-kD cytoplasmic protein, which may play a role in the signal transduction pathway mediated by multiple Wnt proteins. The mechanisms of dishevelled function in Wnt signaling are likely to be conserved among metazoans. [provided by RefSeq, Jul 2008].

Function : Participates in Wnt signaling by binding to the cytoplasmic C-terminus of frizzled family members and transducing the Wnt signal to down-stream effectors. Promotes internalization and degradation of frizzled proteins upon Wnt signaling. Plays a role both in canonical and non-canonical Wnt signaling. Plays a role in the signal transduction pathways mediated by multiple Wnt genes.

Subunit : Interacts through its PDZ domain with the C-terminal regions of VANGL1 and VANGL2. Interacts with Rac. Interacts with ARRB1; the interaction is enhanced by phosphorylation of DVL1. Can form large oligomers (via DIX domain). Interacts (via DIX domain) with DIXDC1 (via DIX domain). Interacts (via DEP domain) with AP2M1 and the AP-2 complex. Interacts with DACT1.

Subcellular Location : Cell membrane; Peripheral membrane protein; Cytoplasmic side. Cytoplasm, cytosol. Cytoplasmic vesicle. Note=Localizes at the cell membrane upon interaction with frizzled family members and promotes their internalization. Localizes to cytoplasmic puncta.

Post-translational modifications : Phosphorylated by CSNK1D.

Similarity : Belongs to the DSH family.

Contains 1 DEP domain.

Contains 1 DIX domain.

Contains 1 PDZ (DHR) domain.

Database links : NCBI Reference Sequence: NP_004413.1

UniProtKB/Swiss-Prot: O14641.1

抗體的鑒定：

1）蓬亂蛋白2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)率可用競爭抑制試驗(yàn)測定。以不同濃度抗原和近似抗原分別做競爭抑制曲線,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。

Integrin alpha E2 peptide 整合素alpha E2抗原Multi-class antibodies規(guī)格： 0.5mg

Anti-phospho-APS(Ser598) 磷酸化APS抗體Multi-class antibodies規(guī)格： 0.1ml

Rhesus antibody Rh Phospho-BTK/BPK(Tyr223) 磷酸化蛋白酪氨酸激酶ATK抗體 規(guī)格 0.1ml

胰蛋白酶工作液(水劑、中度消化) 10ml pH 7.8, 0.05%，免疫組化用

VCAM-1 英文名稱： 內(nèi)皮細(xì)胞粘附分子抗體 0.1ml

DENND2D 英文名稱： DENN域內(nèi)含蛋白2D抗體 0.2ml

Anti-phospho-APS(Ser598) 磷酸化APS抗體Multi-class antibodies規(guī)格： 0.1ml

Anti-OCT1 /FITC 熒光素標(biāo)記陽離子轉(zhuǎn)運(yùn)器1抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rabbit Anti-Goat IgM/HRP 辣根過氧化物酶標(biāo)記兔抗羊IgMMulti-class antibodies規(guī)格： 0.1ml

周期素依賴性激酶5抗體 Anti-CDK5 1ml

Goat Anti-Bovine IgG/Bio 生物素標(biāo)記的羊抗IgG 0.1ml

FGF4 英文名稱： 纖維母細(xì)胞生長因子4抗體 0.1ml

Rhesus antibody Rh PLAP/Phospholipase A2 activator protein 磷脂酶A2激活蛋白抗體 規(guī)格 0.1ml

Rabbit Anti-Goat IgM/HRP 辣根過氧化物酶標(biāo)記兔抗羊IgMMulti-class antibodies規(guī)格： 0.1ml

SASS6 英文名稱： 紡錘體組裝異常蛋白6抗體 0.2ml

EPHA10 英文名稱： 酪酸蛋白激酶受體A10抗體 0.1ml

惡性特異性生長因子抗體 Anti-TSGF 0.1ml

Anti-TRF/FITC 熒光素標(biāo)記抗轉(zhuǎn)鐵蛋白抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh phospho-IRF3(Ser386) 磷酸化干擾素調(diào)節(jié)因子3抗體 規(guī)格 0.1ml

SM-MHC(Cardiac myosin heavy chain) 心肌肌凝蛋白多肽Multi-class antibodies規(guī)格： 0.5mg

U266B1 [U266], 人細(xì)胞

IEC-6 大鼠小腸引窩上皮細(xì)胞

HA Others H1N1 甲型 H1N1 (A/New York/18/2009) HA 人細(xì)胞裂解液 (陽性對(duì)照)

CM-H115人腦動(dòng)脈內(nèi)皮細(xì)胞完全培養(yǎng)基100mL

人*上皮細(xì)胞HREpiC

人細(xì)胞;Hela 人晶狀體上皮細(xì)胞完全培養(yǎng)基 100mL

CL-0016A549(人非小細(xì)胞細(xì)胞)5×106cells/瓶×2

CDH1 Others Rat 大鼠 E-Cadherin / CDH1 / E-cad / CD324 人細(xì)胞裂解液 (陽性對(duì)照)

前脂肪細(xì)胞生長添加物PAGS

WERI-Rb-1細(xì)胞，人視網(wǎng)膜母細(xì)胞瘤 人胚腎細(xì)胞,AD293細(xì)胞 小氣道平滑肌細(xì)胞Many types of cells包裝：5 × 105次方(1ml)

盤羊皮膚細(xì)胞;OAM-S1

SELL Others Mouse 小鼠 SELL / CD62L / L-selectin 人細(xì)胞裂解液 (陽性對(duì)照)

蓬亂蛋白2抗體規(guī)格 U266B1 [U266], 人細(xì)胞

IEC-6 大鼠小腸引窩上皮細(xì)胞

HA Others H1N1 甲型 H1N1 (A/New York/18/2009) HA 人細(xì)胞裂解液 (陽性對(duì)照)

CM-H115人腦動(dòng)脈內(nèi)皮細(xì)胞完全培養(yǎng)基100mL

人*上皮細(xì)胞HREpiC

人細(xì)胞;Hela 人晶狀體上皮細(xì)胞完全培養(yǎng)基 100mL