抗体的生物素化标记实验要点：

1. Ras样蛋白A抗体说明书 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-RALA 

中文名称   Ras样蛋白A抗体说明书 

别    名  Ral a; Ral A protein; RAL; RALA; RALA_HUMAN; RAS like protein A; Ras related protein RalA; Ras-related protein Ral-A.

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, Chicken, Dog, Pig, Cow, Horse, Rabbit 

产品类型   一抗  

研究领域     细胞生物 免疫学 信号转导 G蛋白偶联受体

蛋白分子量  predicted molecular weight: 24kDa

性    状   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human RalA 

亚    型   IgG

纯化方法   affinity purified by Protein A

储 存 液   0.01M PBS, pH 7.4 with 10 mg/ml BSA and 0.1% Sodium azide

Ras样蛋白A抗体说明书 产品应用    WB=1:100-500 ELISA=1:500-1000 IP=1:20-100 IHC-P=1:100-500 IHC-F=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 Multifuntional GTPase involved in a variety of cellular processes including gene expression, cell migration, cell proliferation, oncogenic transformation and membrane trafficking. Accomplishes its multiple functions by interacting with distinct downstream effectors. Acts as a GTP sensor for GTP-dependent exocytosis of dense core vesicles. Plays a role in the early stages of cytokinesis and is required to tether the exocyst to the cytokinetic furrow. The RALA-exocyst complex regulates integrin-dependent membrane raft exocytosis and growth signaling. Key regulator of LPAR1 signaling and competes with ADRBK1 for binding to LPAR1 thus affecting the signaling properties of the receptor. Required for anchorage-independent proliferation of transformed cells.

Function : Multifunctional GTPase involved in a variety of cellular processes including gene expression, cell migration, cell proliferation, oncogenic transformation and membrane trafficking. Accomplishes its multiple functions by interacting with distinct downstream effectors. Acts as a GTP sensor for GTP-dependent exocytosis of dense core vesicles. Plays a role in the early stages of cytokinesis and is required to tether the exocyst to the cytokinetic furrow. The RALA-exocyst complex regulates integrin-dependent membrane raft exocytosis and growth signaling. Key regulator of LPAR1 signaling and competes with ADRBK1 for binding to LPAR1 thus affecting the signaling properties of the receptor. Required for anchorage-independent proliferation of transformed cells.

Subunit : Interacts with RALBP1 via its effector domain. Interacts with EXOC8 and EXOC2. EXOC2 and EXOC8 have overlapping binding sites and compete for RALA binding. Interacts with Clostridium exoenzyme C3. Interacts with RALGPS1. Interacts with LPAR1 and LPAR2. Interacts with ADRBK1 in response to LPAR1 activation. RALA and ADRBK1 mutually inhibit each other's binding to LPAR1.

Subcellular Location : Cell surface. Cell membrane; Lipid-anchor; Cytoplasmic side. Cleavage furrow. Midbody.

Post-translational modifications : Prenylation is essential for membrane localization. The geranylgeranylated form and the farnesylated mutant does not undergo alternative prenylation in response to geranylgeranyltransferase I inhibitors (GGTIs) and farnesyltransferase I inhibitors (FTIs).

Similarity : Belongs to the small GTPase superfamily. Ras family.

Database links : UniProtKB/Swiss-Prot: P11233.1

抗体的鉴定：

1）Ras样蛋白A抗体说明书 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。

MMP-20(matrix metalloproteinase 20) 基质金属蛋白酶20抗原Multi-class antibodies规格： 0.5mg

Anti-Ankyrin G 锚定蛋白G抗体Multi-class antibodies规格： 0.2ml

Rhesus antibody Rh Phospho-Beta-Catenin(Ser33/37) 磷酸化β 连环素蛋白抗体 规格 0.1ml

HRP-Streptavidin工作液 12ml 进口分装

VGF 英文名称： 神经调节肽VGF抗体 0.1ml

DLGAP1 英文名称： PSD95结合蛋白1抗体 0.2ml

Anti-Ankyrin G 锚定蛋白G抗体Multi-class antibodies规格： 0.2ml

Anti-ompF/FITC 荧光素标记小肠耶尔森氏菌膜通道蛋白抗体IgGMulti-class antibodies规格： 0.2ml

Rabbit Anti-human IgA/HRP 辣根过氧化物酶标记兔抗人IgAMulti-class antibodies规格： 0.1ml

软骨糖蛋白39抗体 Anti-CDR/YKL-40 0.1ml

Rabbit Anti-horse IgM/Cy7 Cy7标记的兔抗马IgM 0.1ml

FAM29A 英文名称： FAM29蛋白抗体 0.2ml

Rhesus antibody Rh PLAGL1/ZAC1 多型性腺瘤基因样蛋白1抗体 规格 0.2ml

Rabbit Anti-human IgA/HRP 辣根过氧化物酶标记兔抗人IgAMulti-class antibodies规格： 0.1ml

Sca1 英文名称： T淋巴细胞激活蛋白抗体 0.2ml

LPA1 英文名称： 溶血磷脂酸受体蛋白1抗体 0.1ml

一种新的丝氨酸/苏氨酸激酶家族成员的基因WNK4抗体 Anti-WNK4 0.2ml

Anti-TSHR(NT)/FITC 荧光素标记促甲状腺素受体抗体(N端)IgGMulti-class antibodies规格： 0.2ml

Rhesus antibody Rh phospho-IKK beta(Ser733) 磷酸化KB抑制蛋白激酶β抗体 规格 0.1ml

TRA16 (Testicular orphan nuclear receptor-4 (TR4)-associated protein) 激素受体相关蛋白Multi-class antibodies规格： 0.5mg

HBE, 正常人上皮细胞系

DQSHS1 迪庆绵羊皮肤成纤维样细胞

OMG Others Human 人 OMG / OMGP (aa 1-416) 人细胞裂解液 (阳性对照)

CM-M062小鼠肾小管上皮细胞完全培养基100mL

成纤维细胞培养基FM-acf-prf

EB病毒转化的人B淋巴细胞;KMY0903 新生儿细胞完全培养基 100mL

CL-0068COS-1(非洲绿猴SV40转化的肾细胞)5×106cells/瓶×2

PTPRC Others Mouse 小鼠 CD45 / PTPRC 人细胞裂解液 (阳性对照)

CCCSCK

8910PM细胞，人细胞 小鼠细胞,EMT6细胞 肾小管上皮细胞Many types of cells包装：5 × 105次方(1ml)

Dami(人巨核细胞细胞) 5×106cells/瓶×2

HTEpC Pellet 人类气管上皮细胞团块 > 1 mio.cells 人骨骼肌细胞cDNAHSkMC cDNA

Ras样蛋白A抗体说明书 HBE, 正常人上皮细胞系

DQSHS1 迪庆绵羊皮肤成纤维样细胞

OMG Others Human 人 OMG / OMGP (aa 1-416) 人细胞裂解液 (阳性对照)

CM-M062小鼠肾小管上皮细胞完全培养基100mL

成纤维细胞培养基FM-acf-prf

EB病毒转化的人B淋巴细胞;KMY0903 新生儿细胞完全培养基 100mL