抗體的生物素化標(biāo)記實(shí)驗(yàn)要點(diǎn)：

1. 跨膜蛋白49抗體費(fèi)用 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(lái)篩選最適條件；

3.用NHSB量過(guò)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-TMEM49 

中文名稱   跨膜蛋白49抗體費(fèi)用 

別    名  Vacuole membrane protein 1; DKFZP566I133; TDC1; TMEM49; Transmembrane protein 49; vmp1; VMP1_HUMAN.

濃    度   1mg/1ml

規(guī) 格  0.2ml/200μg

抗體來(lái)源   Rabbit

克隆類型   polyclonal

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

產(chǎn)品類型   一抗  

研究領(lǐng)域     心 細(xì)胞生物 細(xì)胞生物標(biāo)志物 細(xì)胞自噬 細(xì)胞膜蛋白

蛋白分子量  predicted molecular weight: 46kDa

性    狀   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human TMEM49 (361-406aa) 

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

跨膜蛋白49抗體費(fèi)用 產(chǎn)品應(yīng)用    WB=1:100-500 ELISA=1:500-1000 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 Vacuole membrane protein 1 (VMP1)/TMEM49 is a transmembrane protein localized to intracellular vacuoles and was discovered as a protein that promotes vacuole formation in acinar cells associated with acute pancreatitis (1). Over-expression of VMP1 promotes vacuole formation and subsequent cell death (1). Subsequent studies have shown that VMP1 expression is induced by starvation and the mTOR inhibitor, rapamycin, and can trigger autophagy (2). VMP1 is targeted, along with LC3, to autophagosome membranes (2). Knockdown of VMP1 can inhibit autophagosome formation (2). VMP1 interacts with Beclin-1, a key autophagy protein that activates the Class III PI3 kinase Vps34, which is regulated by a large network of associated proteins (3). VMP1 functions in the degradation and clearance of zymogen-containing vacuoles during experimental pancreatitis (4). During this process, VMP1 interacts with the ubiquitin protease USP9X, suggesting a possible functional link between the molecular machinery of autophagy and the ubiquitin pathway. Orthologues of VMP1 have been reported in C. elegans (known as EPG-3), Drosophila (known as TANGO-5), and Dictyostelium, and have been shown to play a role in membrane trafficking, orgenelle organization, and autophagy (5-7).

Function : Stress-induced protein that, when overexpressed, promotes formation of intracellular vacuoles followed by cell death. May be involved in the cytoplasmic vacuolization of acinar cells during the early stage of acute pancreatitis. Plays a role in the initial stages of the autophagic process through its interaction with BECN1 (By similarity). Involved in cell-cell adhesion. Plays an essential role in formation of cell junctions.Sequence similarities: Belongs to the VMP1 family.

Subunit : Belongs to the VMP1 family.

Subcellular Location : Endoplasmic reticulum-Golgi intermediate compartment membrane. Cell membrane. Vacuole membrane. Endoplasmic reticulum.

Similarity : Belongs to the VMP1 family.

Database links : UniProtKB/Swiss-Prot: Q96GC9.1

抗體的鑒定：

1）跨膜蛋白49抗體費(fèi)用 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(lái)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(lái)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(wú)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。

OAT-3(Organic anion transporter 3) 陰離子轉(zhuǎn)運(yùn)蛋白-3(抗原)Multi-class antibodies規(guī)格： 0.5mg

Anti-CD45 CD45單克隆抗體Multi-class antibodies規(guī)格： 0.1ml

Rhesus antibody Rh P53 protein(wt-p53) 抑制基因P53蛋白抗體(野生型P53) 規(guī)格 0.1ml

ICAM1 Kit Rat 大鼠 ICAM-1 / CD54 ELISA配對(duì)抗體 ELISA

TSPAN5 英文名稱： 四分子交聯(lián)體5抗體(四旋蛋白) 0.2ml

C1RL 英文名稱： 補(bǔ)體C1RL鏈多肽抗體 0.2ml

Anti-CD45 CD45單克隆抗體Multi-class antibodies規(guī)格： 0.1ml

Nephrin Protein 去氧Multi-class antibodies規(guī)格： 0.5mg

Anti-APEX1 多功能DNA修復(fù)酶抗體Multi-class antibodies規(guī)格： 0.1ml

Rhesus antibody Rh phospho-BACE1(Ser498) 磷酸化β分泌酶抗體 規(guī)格 0.1ml

品名 規(guī)格 備注

Vitamin D Binding protein 英文名稱： 維生素D結(jié)合蛋白抗體 0.2ml

DAP12 英文名稱： 自然殺傷激活受體相關(guān)蛋白DAP12抗體 0.1ml

Anti-APEX1 多功能DNA修復(fù)酶抗體Multi-class antibodies規(guī)格： 0.1ml

Anti-TTF-1/FITC 熒光素標(biāo)記甲狀腺核轉(zhuǎn)錄因子-1/甲狀腺特異增強(qiáng)子結(jié)合蛋白抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Trk B 神經(jīng)生長(zhǎng)因子受體的一種(抗原)Multi-class antibodies規(guī)格： 0.5mg

CASPASE3凋亡抑制劑 Anti-Z-DEVD-FMK 0.2ml

Smoothened 英文名稱： 跨膜蛋白Smoothened抗體 0.1ml

Epiphycan 英文名稱： 軟骨素蛋白多糖3抗體 0.2ml

Rhesus antibody Rh phospho-IKK alpha (Tyr463) 磷酸化KB抑制蛋白激酶α抗體 規(guī)格 0.1ml

Trk B 神經(jīng)生長(zhǎng)因子受體的一種(抗原)Multi-class antibodies規(guī)格： 0.5mg

大鼠硫氧還蛋白還原酶(xR)ELISA試劑盒 ，英文名： xR ELISA Kit

人基質(zhì)金屬蛋白酶抑制因子1(TIMP-1)ELISA檢測(cè)試劑盒Humaissueinhibitorsofmetalloproteinase1,TIMP-1ELISAKit 96T/48T

鴨白介素2(IL-2)免疫試劑盒 Duck Ierleukin 2,IL-2 ELISA kit

CLIAKitforp53(Rabbitp53/tumorprotein)ELISAKit兔p53規(guī)格：48T/96T

水中隱孢子蟲(Cryptosporidium)基因熒光定量檢測(cè)試劑盒20次

ELISAKitLTC4人白三烯C4規(guī)格：48T/96T

大鼠補(bǔ)體成分5(C5)ELISA試劑盒 ，英文名： C5 ELISA Kit

Human alpha N is acyl Glucosaminidase (NAG) ELISA Kit 人αN已酰氨基葡糖苷酶(αNAG)ELISA試劑盒

RatIerleukin27,IL-27ELISAKit 大鼠白介素27(IL-27)ELISA試劑盒 96T/48T 進(jìn)口分裝

CLIAKitforCP(HumancicaicialPemphigoid)ELISAKit人性天皰瘡抗體規(guī)格：48T/96T

細(xì)胞合成酶總活性比色法定量檢測(cè)試劑盒20次

Ratcoicoopieleasinghormone,CRHELISAKit大鼠促腎上皮質(zhì)激素釋放激素(CRH)ELISA試劑盒規(guī)格：96T/48T

跨膜蛋白49抗體費(fèi)用 大鼠蛋酸酶(PP)ELISA 試劑盒 96T/48T 試劑盒 組裝/原裝

人內(nèi)肽2(EM-2)免疫試劑盒 Human Endomorphin-2,EM-2 ELISA Kit

MouseTestoterone,TELISAKit小鼠睪同(T)ELISA試劑盒規(guī)格：96T/48T

二(MDA)終點(diǎn)比色法定量檢測(cè)試劑盒50次

Humanplatelet-derivedgrowthfactor,PDGFELISA試劑盒人血小板衍生生長(zhǎng)因子(PDGF)ELISA試劑盒規(guī)格：96T/48T

HumanSurfaceMembraneImmunoglobulin，SmIgELISAKit人細(xì)胞膜表面免疫球蛋白(SmIg)ELISA試劑盒規(guī)格：96T/48T