抗体的生物素化标记实验要点：

1. 跨膜蛋白49抗体费用 如在反应混合液中有叠氮钠或游离氨基存在,会抑制标记反应。因此,蛋白质在反应前要对 0.1mol/L碳酸氢钠缓冲液或0.5mol/L硼酸缓冲液充分透析；

2.所用的NHSB及待生物素化蛋白质之间的分子比按蛋白质表面的ε-氨基的密度会有所不同,选择不当则影响标记的效率,应先用几个不同的分子比来筛选最适条件；

3.用NHSB量过量也是不利的,抗原的结合位点可能因此被封闭,导致抗体失活；

4.由于抗体的氨基不易接近可能造成生物素化不足,此时可加入去污剂如 Triton x-100, Tween20等；

5.当游离ε-氨基(赖氨酸残基的氨基)存在于抗体的抗原结合位点时,或位于酶的催化位点时,生物素化会降低或损伤抗体蛋白的结合力或活性；

6.生物素还可能与不同的功能基团,如羰基、氨基、巯基、异咪唑基及苯酚基,也可与糖基共价结合；

7.交联反应后,应充分透析,否则,残余的生物素会对生物素化抗体与亲和素的结合产生竞争作用；

8.在细胞的荧光标记实验中,中和亲和素的本底低,但由于链霉亲和素含有少量正电荷,故对某些细胞可导致高本底。

产品订购信息：
英文名称  Anti-TMEM49 

中文名称   跨膜蛋白49抗体费用 

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

浓    度   1mg/1ml

规 格  0.2ml/200μg

抗体来源   Rabbit

克隆类型   polyclonal

交叉反应   Human, Mouse, Rat, Pig, Horse, Rabbit 

产品类型   一抗  

研究领域     心 细胞生物 细胞生物标志物 细胞自噬 细胞膜蛋白

蛋白分子量  predicted molecular weight: 46kDa

性    状   Lyophilized or Liquid

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

亚    型   IgG

纯化方法   affinity purified by Protein A

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

跨膜蛋白49抗体费用 产品应用    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

（石蜡切片需做抗原修复） 

 not yet tested in other applications.

 optimal dilutions/concentrations should be determined by the end user.  

保存条件  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. 

产品介绍 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

抗体的鉴定：

1）跨膜蛋白49抗体费用 抗体的效价鉴定：不管是用于诊断还是用于,制备抗体的目的都是要求较高效价。不同的抗原制备的抗体,要求的效价不一。鉴定效价的方法很多,包括有试管凝集反应,琼脂扩散试验,酶联免疫吸附试验等。常用的抗原所制备的抗体一般都有约成的鉴定效价的方法,以资比较。如制备抗抗体的效价,一般就采用琼脂扩散试验来鉴定。

2）抗体的特异性鉴定：抗体的特异性是指与相应抗原或近似抗原物质的识别能力。抗体的特异性高,它的识别能力就强。衡量特异性通常以交叉反应率来表示。交叉反应率可用竞争抑制试验测定。以不同浓度抗原和近似抗原分别做竞争抑制曲线,计算各自的结合率,求出各自在IC50时的浓度,并按公式计算交叉反应率。 

如果所用抗原浓度IC50浓度为pg/管,而一些近似抗原物质的IC50浓度几乎是无穷大时,表示这一抗血清与其他抗原物质的交叉反应率近似为0,即该血清的特异性较好。

3）抗体亲和力：是指抗体和抗原结合的牢固程度。亲和力的高低是由抗原分子的大小,抗体分子的结合位点与抗原决定簇之间立体构型的合适度决定的。有助于维持抗原抗体复合物稳定的分子间力有氢键,疏水键,侧链相反电荷基因的库仑力,范德华力和空间斥力。亲和力常以亲和常数K表示,K的单位是L/mol。抗体亲和力的测定对抗体的筛选,确定抗体的用途,验证抗体的均一性等均有重要意义。

OAT-3(Organic anion transporter 3) 阴离子转运蛋白-3(抗原)Multi-class antibodies规格： 0.5mg

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Rhesus antibody Rh P53 protein(wt-p53) 抑制基因P53蛋白抗体(野生型P53) 规格 0.1ml

ICAM1 Kit Rat 大鼠 ICAM-1 / CD54 ELISA配对抗体 ELISA

TSPAN5 英文名称： 四分子交联体5抗体(四旋蛋白) 0.2ml

C1RL 英文名称： 补体C1RL链多肽抗体 0.2ml

Anti-CD45 CD45单克隆抗体Multi-class antibodies规格： 0.1ml

Nephrin Protein 去氧Multi-class antibodies规格： 0.5mg

Anti-APEX1 多功能DNA修复酶抗体Multi-class antibodies规格： 0.1ml

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品名 规格 备注

Vitamin D Binding protein 英文名称： 维生素D结合蛋白抗体 0.2ml

DAP12 英文名称： 自然杀伤激活受体相关蛋白DAP12抗体 0.1ml

Anti-APEX1 多功能DNA修复酶抗体Multi-class antibodies规格： 0.1ml

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Trk B 神经生长因子受体的一种(抗原)Multi-class antibodies规格： 0.5mg

CASPASE3凋亡抑制剂 Anti-Z-DEVD-FMK 0.2ml

Smoothened 英文名称： 跨膜蛋白Smoothened抗体 0.1ml

Epiphycan 英文名称： 软骨素蛋白多糖3抗体 0.2ml

Rhesus antibody Rh phospho-IKK alpha (Tyr463) 磷酸化KB抑制蛋白激酶α抗体 规格 0.1ml

Trk B 神经生长因子受体的一种(抗原)Multi-class antibodies规格： 0.5mg

大鼠硫氧还蛋白还原酶(xR)ELISA试剂盒 ，英文名： xR ELISA Kit

人基质金属蛋白酶抑制因子1(TIMP-1)ELISA检测试剂盒Humaissueinhibitorsofmetalloproteinase1,TIMP-1ELISAKit 96T/48T

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水中隐孢子虫(Cryptosporidium)基因荧光定量检测试剂盒20次

ELISAKitLTC4人白三烯C4规格：48T/96T

大鼠补体成分5(C5)ELISA试剂盒 ，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试剂盒

RatIerleukin27,IL-27ELISAKit 大鼠白介素27(IL-27)ELISA试剂盒 96T/48T 进口分装

CLIAKitforCP(HumancicaicialPemphigoid)ELISAKit人性天疱疮抗体规格：48T/96T

细胞合成酶总活性比色法定量检测试剂盒20次

Ratcoicoopieleasinghormone,CRHELISAKit大鼠促肾上皮质激素释放激素(CRH)ELISA试剂盒规格：96T/48T

跨膜蛋白49抗体费用 大鼠蛋酸酶(PP)ELISA 试剂盒 96T/48T 试剂盒 组装/原装

人内肽2(EM-2)免疫试剂盒 Human Endomorphin-2,EM-2 ELISA Kit

MouseTestoterone,TELISAKit小鼠睾同(T)ELISA试剂盒规格：96T/48T

二(MDA)终点比色法定量检测试剂盒50次

Humanplatelet-derivedgrowthfactor,PDGFELISA试剂盒人血小板衍生生长因子(PDGF)ELISA试剂盒规格：96T/48T

HumanSurfaceMembraneImmunoglobulin，SmIgELISAKit人细胞膜表面免疫球蛋白(SmIg)ELISA试剂盒规格：96T/48T