抗體的生物素化標(biāo)記實(shí)驗(yàn)要點(diǎn)：

1. 三磷酸腺苷結(jié)合轉(zhuǎn)運(yùn)蛋白G超家族成員8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。

產(chǎn)品訂購(gòu)信息：
英文名稱  Anti-STSL/ABCG8 

中文名稱   三磷酸腺苷結(jié)合轉(zhuǎn)運(yùn)蛋白G超家族成員8抗體說明書 

別    名  ATP binding cassette sub family G (WHITE) member 8 (sterolin 2); ATP binding cassette sub family G member 8; MGC142217; Sterolin 2; STSL; ABCG8_HUMAN.

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, Dog, Pig, Cow, Horse, Rabbit 

產(chǎn)品類型   一抗  

研究領(lǐng)域    

蛋白分子量  predicted molecular weight: 76kDa

性    狀   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human ABCG8 

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

三磷酸腺苷結(jié)合轉(zhuǎn)運(yùn)蛋白G超家族成員8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 Flow-Cyt=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 The protein encoded by this gene is a member of the superfamily of ATP-binding cassette (ABC) transporters. ABC proteins transport various molecules across extra- and intra-cellular membranes. ABC genes are divided into seven distinct subfamilies (ABC1, MDR/TAP, MRP, ALD, OABP, GCN20, White). This protein is a member of the White subfamily. The protein encoded by this gene functions to exclude non-cholesterol sterol entry at the intestinal level, promote excretion of cholesterol and sterols into bile, and to facilitate transport of sterols back into the intestinal lumen. It is expressed in a tissue-specific manner in the liver, intestine, and gallbladder. This gene is tandemly arrayed on chromosome 2, in a head-to-head orientation with family member ABCG5. Mutations in this gene may contribute to sterol accumulation and atherosclerosis, and have been observed in patients with sitosterolemia. [provided by RefSeq, Jul 2008].

Function : Transporter that appears to play an indispensable role in the selective transport of the dietary cholesterol in and out of the enterocytes and in the selective sterol excretion by the liver into bile.

Subunit : May form heterodimers with ABCG5 or be tightly coupled to ABCG5 along a pathway regulating diatery-sterol absorption and excretion.

Subcellular Location : Membrane; Multi-pass membrane protein (Probable).

Tissue Specificity : Strongly expressed in the liver, lower levels in the small intestine and colon. Detectable in a wide variety of human tissues.

DISEASE : Genetic variations in ABCG8 can be associated with susceptibility to gallbladder disease type 4 (GBD4) [MIM:611465]. With an overall prevalence of 10-20%, gallstone disease (cholelithiasis) represents one of the most frequent and economically relevant health problems of industrialized countries.

Defects in ABCG8 are a cause of sitosterolemia (STSL) [MIM:210250]; also known as phytosterolemia or shellfish sterolemia. It is a rare autosomal recessive disorder characterized by increased intestinal absorption of all sterols including cholesterol, plant and shellfish sterols, and decreased biliary excretion of dietary sterols into bile. Sitosterolemia patients have hypercholesterolemia, very high levels of plant sterols in the plasma, and frequently develop tendon and tuberous xanthomas, accelerated atherosclerosis and premature coronary artery disease.

Similarity : Belongs to the ABC transporter superfamily. ABCG family. Eye pigment precursor importer (TC 3.A.1.204) subfamily.

Contains 1 ABC transmembrane type-2 domain.

Contains 1 ABC transporter domain.

Database links : UniProtKB/Swiss-Prot: Q9H221.1

抗體的鑒定：

1）三磷酸腺苷結(jié)合轉(zhuǎn)運(yùn)蛋白G超家族成員8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。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(kù)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-UMOD/FITC 熒光素標(biāo)記UMOD抗體IgGMulti-class antibodies規(guī)格： 0.2ml

CXCL10/IP-10 CXCL10趨化因子10/干擾素誘導(dǎo)蛋白10抗原Multi-class antibodies規(guī)格： 0.5mg

組織蛋白酶H抗體 Anti-cath-H/CH 0.1ml

Sec8 英文名稱： 胞吐分泌蛋白Sec8抗體 0.2ml

EDRF1 英文名稱： 紅細(xì)胞分化相關(guān)因子1抗體 0.2ml

Rhesus antibody Rh Phospho-Histone H3 (Thr11) 磷酸化組蛋白H3抗體 規(guī)格 0.1ml

CXCL10/IP-10 CXCL10趨化因子10/干擾素誘導(dǎo)蛋白10抗原Multi-class antibodies規(guī)格： 0.5mg

Anti-Phospho-FAK (Tyr925) /FITC 熒光素標(biāo)記兔抗人、大、小鼠磷酸化粘著斑激酶抗體IgGMulti-class antibodies規(guī)格： 0.2ml

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

角質(zhì)形成細(xì)胞生長(zhǎng)因子受體/纖維母細(xì)胞生長(zhǎng)因子-7抗體 Anti- KGF/FGF-7 0.1ml

Rabbit Anti-Goat IgG/PE PE標(biāo)記的兔抗羊IgG 0.1ml

FOXRED1 英文名稱： 單跨膜蛋白FOXRED1抗體 0.2ml

Rhesus antibody Rh Phospho-Syk (Ty*3) 磷酸化非受體型酪氨酸蛋白激酶抗體 規(guī)格 0.1ml

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

Resistin 抵抗素抗原Multi-class antibodies規(guī)格： 0.5mg

Anti-phospho-Bax(Ser184) 磷酸化Bax抗體Multi-class antibodies規(guī)格： 0.1ml

Rhesus antibody Rh PHLP 抑制基因PHLPP抗體 規(guī)格 0.2ml

Keratin10 濃縮液 0.1ml 進(jìn)口分裝

UVRAG 英文名稱： 抑制基因UVRAG抗體 0.2ml

Chorein/VPS13A 英文名稱： 液泡蛋白分選蛋白VPS13A抗體 0.1ml

Anti-phospho-Bax(Ser184) 磷酸化Bax抗體Multi-class antibodies規(guī)格： 0.1ml

IL1B Protein Cynomolgus 重組食蟹猴 IL-1 beta / IL1B 蛋白

小鼠肺大動(dòng)脈平滑肌細(xì)胞完全培養(yǎng)基 100mL

MFI2 Others Mouse 小鼠 MFI2 / CD228 / melanoansferrin 人細(xì)胞裂解液 (陽性對(duì)照)

C3H/10T1/2, Clone 8小鼠胚胎成纖維細(xì)胞 C3H/10T1/2, Clone 8 mouse embryonic fibroblast MEM培養(yǎng)基(GIBCO)+10%FBS

CL-0345HB611(人細(xì)胞)5×106cells/瓶×2

NCI-H520(人肺鱗癌細(xì)胞) 5×106cells/瓶×2 T/G HA-VSMC(人主動(dòng)脈平滑肌細(xì)胞)

CM-R037大鼠小腸隱窩上皮細(xì)胞完全培養(yǎng)基100mL

IGFBP5 Others Cynomolgus 食蟹猴 IGFBP5 / IGFBP-5 人細(xì)胞裂解液 (陽性對(duì)照)

人脂肪間充質(zhì)干細(xì)胞HMSC-ad

SK-MES-1細(xì)胞，人細(xì)胞 人膀胱鱗癌細(xì)胞,ScaBER細(xì)胞 人前脂肪細(xì)胞-皮下HPA-s

水貂肺上皮細(xì)胞;Mv.1.Lu(NBL-7)

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

三磷酸腺苷結(jié)合轉(zhuǎn)運(yùn)蛋白G超家族成員8抗體說明書 IL1B Protein Cynomolgus 重組食蟹猴 IL-1 beta / IL1B 蛋白

小鼠肺大動(dòng)脈平滑肌細(xì)胞完全培養(yǎng)基 100mL

MFI2 Others Mouse 小鼠 MFI2 / CD228 / melanoansferrin 人細(xì)胞裂解液 (陽性對(duì)照)

C3H/10T1/2, Clone 8小鼠胚胎成纖維細(xì)胞 C3H/10T1/2, Clone 8 mouse embryonic fibroblast MEM培養(yǎng)基(GIBCO)+10%FBS

CL-0345HB611(人細(xì)胞)5×106cells/瓶×2

NCI-H520(人肺鱗癌細(xì)胞) 5×106cells/瓶×2 T/G HA-VSMC(人主動(dòng)脈平滑肌細(xì)胞)