产品订购信息：
英文名称  Anti-RACGAP1 

中文名称   Rho GTP酶激活蛋白GAP抗体说明书 

别    名  GAP; Gap1; GTPase activating protein; HsCYK 4; HsCYK4; ID GAP; KIAA1478; MgcRacGAP; Rac GTPase activating protein 1; RACGAP 1; RGAP1_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, Dog, Cow 

产品类型   一抗  

研究领域    细胞生物 信号转导 转录调节因子 细胞分化 G蛋白信号

蛋白分子量  predicted molecular weight: 71kDa

性    状   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from hu RACGAP1 

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

Rho GTP酶激活蛋白GAP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 Rho GTPases control a variety of cellular processes. There are 3 subtypes of Rho GTPases in the Ras superfamily of small G proteins: RHO, RAC and CDC42. GTPase-activating proteins (GAPs) bind activated forms of Rho GTPases and stimulate GTP hydrolysis. Through this catalytic function, Rho GAPs negatively regulate Rho-mediated signals. GAPs may also serve as effector molecules and play a role in signaling downstream of Rho and other Ras-like GTPases.

Function : Component of the centralspindlin complex that serves asa microtubule-dependent and Rho-mediated signaling required for themyosin contractile ring formation during the cell cyclecytokinesis. Plays key roles in controlling cell growth anddifferentiation of hematopoietic cells through mechanisms otherthan regulating Rac GTPase activity. Also involved in theregulation of growth-related processes in adipocytes and myoblasts.May be involved in regulating spermatogenesis and in the RACGAP1pathway in neuronal proliferation. Shows strong GAP (GTPaseactivation) activity towards CDC42 and RAC1 and less towards RHOA.Essential for the early stages of embryogenesis. May play a role inregulating cortical activity through RHOA during cytokinesis. Mayparticipate in the regulation of sulfate transport in male germcells.

Subunit : Heterotetramer of two molecules each of RACGAP1 andKIF23. Found in the centralspindlin complex composed of RACGAP1 andKIF23. Associates with alpha-, beta- and gamma-tubulin andmicrotubules. Interacts via its Rho-GAP domain with RND2.Associates with AURKB during M phase. Interacts via its Rho-GAPdomain and basic region with PRC1. The interaction with PRC1inhibits its GAP activity towards CDC42 in vitro, which may berequired for maintaining normal spindle morphology. Interacts withSLC26A8 via its N-terminus. Interacts with RAB11FIP3. Interactswith ECT2; the interaction is direct, occurs at anaphase and duringcytokinesis in a microtubule-dependent manner and is enhanced byphosphorylation by PLK1. Interacts with KIF23; the interaction isdirect.

Subcellular Location : ucleus. Cytoplasm. Cytoplasm, cytoskeleton,spindle. Cytoplasmic vesicle, secretory vesicle, acrosome. Cleavagefurrow. Midbody. Note=Colocalizes with RND2 in Golgi-derivedproacrosomal vesicles and the acrosome (By similarity). Duringinterphase, localized to the nucleus and cytoplasm along withmicrotubules, in anaphase, is redistributed to the central spindleand, in telophase and cytokinesis, to the midbody. Colocalizes withRHOA at the myosin contractile ring during cytokinesis. Colocalizeswith ECT2 to the mitotic spindles during anaphase/metaphase, thecleavage furrow during telophase and at the midbody at the end ofcytokinesis. Colocalizes wit Cdc42 to spindle microtubules fromprometaphase to telophase.

Tissue Specificity : Highly expressed in testis, thymus andplacenta. Expressed at lower levels in spleen and peripheral bloodlymphocytes. In testis, expression is restricted to germ cells withthe highest levels of expression found in spermatocytes. Expressionis regulated in a cell cycle-dependent manner and peaks during G2/Mphase.

Post-translational modifications : Phosphorylated at multiple sites in the midbody duringcytokinesis. Phosphorylation by AURKB on Ser-387 at the midbody is,at least in part, responsible for exerting its latent GAP activitytowards RhoA. Phosphorylation on multiple serine residues by PLK1enhances its association with ECT2 and is critical for cleavagefurrow formation.

Similarity : Contains 1 phorbol-ester/DAG-type zinc finger.

Contains 1 Rho-GAP domain.

Database links : UniProtKB/Swiss-Prot: Q9H0H5.1

EpCAM(Epithelial cell adhesion molecule) 上皮细胞粘附分子/表皮细胞粘附分子(多肽)Multi-class antibodies规格： 0.5mg

ATP合成酶H+转运线粒体F0复合体亚基F6抗体 Anti-ATP5j 0.1ml

Rhesus antibody Rh phospho-EGFR (Tyr869) 磷酸化表皮生长因子受体抗体 规格 0.1ml

双抗 70ml Gibco

Zonula occludens protein 3 英文名称： 紧密粘连蛋白ZO-3抗体 0.1ml

DOPA Decarboxylase 英文名称： 多巴胺脱羧酶抗体 0.1ml

ATP合成酶H+转运线粒体F0复合体亚基F6抗体 Anti-ATP5j 0.1ml

phospho-FAK(pSer732)(phospho-Focal adhesion kinase) 磷酸化粘着斑激酶抗原Multi-class antibodies规格： 0.5mg

细胞死亡调解子抗体 Anti-Bim 0.1ml

Rhesus antibody Rh phospho-ECT2 (Thr790) 磷酸化上皮细胞癌转化蛋白2抗体 规格 0.1ml

暗室灯 个

ZNF148 英文名称： 锌指蛋白148抗体 0.2ml

DAPL1 英文名称： 凋亡相关蛋白激酶样蛋白1抗体 0.2ml

细胞死亡调解子抗体 Anti-Bim 0.1ml

ZBTB25 英文名称： 锌指蛋白46抗体 0.2ml

DAP1 英文名称： 死亡相关蛋白1抗体 0.2ml

Artemin抗体 Anti-ART 0.1ml

Anti-CD8/PE 荧光素藻红蛋白标记兔抗人、大、小鼠CD8抗体IgGMulti-class antibodies规格： 0.2ml

Rhesus antibody Rh phospho-EIF4EBP1(Ser111) 磷酸化eIF4E结合蛋白抗体 规格 0.1ml

EGF(Epidermal growth factor)rat 重组表皮生长因子抗原(大鼠)Multi-class antibodies规格： 0.5mg

FASLG Protein Human 重组人 Fas Ligand / FASLG / CD95L 蛋白 (His 标签)

CNE-2 低分化细胞

ICOS Others Mouse 小鼠 ICOS / AILIM / CD278 人细胞裂解液 (阳性对照)

BxPC-3人原位*腺癌细胞 BxPC-3 in situ pancreatic adenocarcinoma cells 1640+10% FBS

小鼠细胞(分泌促生长激素分泌激素);AtT-20

人成纤维细胞( HCF) ( 5×105 ) GC-1, 鼠精原细胞 Mouse

CL-0456TM4(正常小鼠Sertoli细胞)5×106cells/瓶×2

SIGIRR Others Human 人 SIGIRR / TIR8 人细胞裂解液 (阳性对照)

人膀胱基质成纤维细胞RNAHBdSF miRNA5 μg

T24细胞，人膀胱移行细胞癌细胞 大鼠细胞(wistar 大鼠),CBRH7919细胞 RN-c, 大鼠皮质神经元

Ishikawa(人细胞) 5×106cells/瓶×2

hPC-PL-c 人类胎盘周细胞(hPC-PL) 500,000cells 小鼠巨噬细胞MMa

Rho GTP酶激活蛋白GAP抗体说明书 FASLG Protein Human 重组人 Fas Ligand / FASLG / CD95L 蛋白 (His 标签)

CNE-2 低分化细胞

ICOS Others Mouse 小鼠 ICOS / AILIM / CD278 人细胞裂解液 (阳性对照)

BxPC-3人原位*腺癌细胞 BxPC-3 in situ pancreatic adenocarcinoma cells 1640+10% FBS

小鼠细胞(分泌促生长激素分泌激素);AtT-20

人成纤维细胞( HCF) ( 5×105 ) GC-1, 鼠精原细胞 Mouse

抗体的生物素化标记实验要点：

1. Rho GTP酶激活蛋白GAP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。

抗体的鉴定：

1）Rho GTP酶激活蛋白GAP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。