產(chǎn)品訂購信息：
英文名稱  Anti-RACGAP1 

中文名稱   Rho GTP酶激活蛋白GAP抗體說明書 

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

產(chǎn)品類型   一抗  

研究領(lǐng)域    細(xì)胞生物 信號轉(zhuǎn)導(dǎo) 轉(zhuǎn)錄調(diào)節(jié)因子 細(xì)胞分化 G蛋白信號

蛋白分子量  predicted molecular weight: 71kDa

性    狀   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from hu RACGAP1 

亞    型   IgG

純化方法   affinity purified by Protein A

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

Rho GTP酶激活蛋白GAP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 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 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) 上皮細(xì)胞粘附分子/表皮細(xì)胞粘附分子(多肽)Multi-class antibodies規(guī)格： 0.5mg

ATP合成酶H+轉(zhuǎn)運(yùn)線粒體F0復(fù)合體亞基F6抗體 Anti-ATP5j 0.1ml

Rhesus antibody Rh phospho-EGFR (Tyr869) 磷酸化表皮生長因子受體抗體 規(guī)格 0.1ml

雙抗 70ml Gibco

Zonula occludens protein 3 英文名稱： 緊密粘連蛋白ZO-3抗體 0.1ml

DOPA Decarboxylase 英文名稱： 多巴胺脫羧酶抗體 0.1ml

ATP合成酶H+轉(zhuǎn)運(yùn)線粒體F0復(fù)合體亞基F6抗體 Anti-ATP5j 0.1ml

phospho-FAK(pSer732)(phospho-Focal adhesion kinase) 磷酸化粘著斑激酶抗原Multi-class antibodies規(guī)格： 0.5mg

細(xì)胞死亡調(diào)解子抗體 Anti-Bim 0.1ml

Rhesus antibody Rh phospho-ECT2 (Thr790) 磷酸化上皮細(xì)胞癌轉(zhuǎn)化蛋白2抗體 規(guī)格 0.1ml

暗室燈 個(gè)

ZNF148 英文名稱： 鋅指蛋白148抗體 0.2ml

DAPL1 英文名稱： 凋亡相關(guān)蛋白激酶樣蛋白1抗體 0.2ml

細(xì)胞死亡調(diào)解子抗體 Anti-Bim 0.1ml

ZBTB25 英文名稱： 鋅指蛋白46抗體 0.2ml

DAP1 英文名稱： 死亡相關(guān)蛋白1抗體 0.2ml

Artemin抗體 Anti-ART 0.1ml

Anti-CD8/PE 熒光素藻紅蛋白標(biāo)記兔抗人、大、小鼠CD8抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh phospho-EIF4EBP1(Ser111) 磷酸化eIF4E結(jié)合蛋白抗體 規(guī)格 0.1ml

EGF(Epidermal growth factor)rat 重組表皮生長因子抗原(大鼠)Multi-class antibodies規(guī)格： 0.5mg

FASLG Protein Human 重組人 Fas Ligand / FASLG / CD95L 蛋白 (His 標(biāo)簽)

CNE-2 低分化細(xì)胞

ICOS Others Mouse 小鼠 ICOS / AILIM / CD278 人細(xì)胞裂解液 (陽性對照)

BxPC-3人原位*腺癌細(xì)胞 BxPC-3 in situ pancreatic adenocarcinoma cells 1640+10% FBS

小鼠細(xì)胞(分泌促生長激素分泌激素);AtT-20

人成纖維細(xì)胞( HCF) ( 5×105 ) GC-1, 鼠精原細(xì)胞 Mouse

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

SIGIRR Others Human 人 SIGIRR / TIR8 人細(xì)胞裂解液 (陽性對照)

人膀胱基質(zhì)成纖維細(xì)胞RNAHBdSF miRNA5 μg

T24細(xì)胞，人膀胱移行細(xì)胞癌細(xì)胞 大鼠細(xì)胞(wistar 大鼠),CBRH7919細(xì)胞 RN-c, 大鼠皮質(zhì)神經(jīng)元

Ishikawa(人細(xì)胞) 5×106cells/瓶×2

hPC-PL-c 人類胎盤周細(xì)胞(hPC-PL) 500,000cells 小鼠巨噬細(xì)胞MMa

Rho GTP酶激活蛋白GAP抗體說明書 FASLG Protein Human 重組人 Fas Ligand / FASLG / CD95L 蛋白 (His 標(biāo)簽)

CNE-2 低分化細(xì)胞

ICOS Others Mouse 小鼠 ICOS / AILIM / CD278 人細(xì)胞裂解液 (陽性對照)

BxPC-3人原位*腺癌細(xì)胞 BxPC-3 in situ pancreatic adenocarcinoma cells 1640+10% FBS

小鼠細(xì)胞(分泌促生長激素分泌激素);AtT-20

人成纖維細(xì)胞( HCF) ( 5×105 ) GC-1, 鼠精原細(xì)胞 Mouse

抗體的生物素化標(biāo)記實(shí)驗(yàn)要點(diǎn)：

1. Rho GTP酶激活蛋白GAP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,會抑制標(biāo)記反應(yīng)。因此,蛋白質(zhì)在反應(yīng)前要對 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,殘余的生物素會對生物素化抗體與親和素的結(jié)合產(chǎn)生競爭作用；

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,故對某些細(xì)胞可導(dǎo)致高本底。

抗體的鑒定：

1）Rho GTP酶激活蛋白GAP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)的識別能力。抗體的特異性高,它的識別能力就強(qiáng)。衡量特異性通常以交叉反應(yīng)率來表示。交叉反應(yīng)率可用競爭抑制試驗(yàn)測定。以不同濃度抗原和近似抗原分別做競爭抑制曲線,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。抗體親和力的測定對抗體的篩選,確定抗體的用途,驗(yàn)證抗體的均一性等均有重要意義。