抗體的生物素化標(biāo)記實(shí)驗(yàn)要點(diǎn)：

1. Ras樣蛋白A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(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(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)品訂購信息：
英文名稱  Anti-RALA 

中文名稱   Ras樣蛋白A抗體說明書 

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

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

產(chǎn)品類型   一抗  

研究領(lǐng)域     細(xì)胞生物 免疫學(xué) 信號(hào)轉(zhuǎn)導(dǎo) G蛋白偶聯(lián)受體

蛋白分子量  predicted molecular weight: 24kDa

性    狀   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human RalA 

亞    型   IgG

純化方法   affinity purified by Protein A

儲(chǔ) 存 液   0.01M PBS, pH 7.4 with 10 mg/ml BSA and 0.1% Sodium azide

Ras樣蛋白A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 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

抗體的鑒定：

1）Ras樣蛋白A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(shí)別能力?？贵w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?？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。

MMP-20(matrix metalloproteinase 20) 基質(zhì)金屬蛋白酶20抗原Multi-class antibodies規(guī)格： 0.5mg

Anti-Ankyrin G 錨定蛋白G抗體Multi-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh Phospho-Beta-Catenin(Ser33/37) 磷酸化β 連環(huán)素蛋白抗體 規(guī)格 0.1ml

HRP-Streptavidin工作液 12ml 進(jìn)口分裝

VGF 英文名稱： 神經(jīng)調(diào)節(jié)肽VGF抗體 0.1ml

DLGAP1 英文名稱： PSD95結(jié)合蛋白1抗體 0.2ml

Anti-Ankyrin G 錨定蛋白G抗體Multi-class antibodies規(guī)格： 0.2ml

Anti-ompF/FITC 熒光素標(biāo)記小腸耶爾森氏菌膜通道蛋白抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rabbit Anti-human IgA/HRP 辣根過氧化物酶標(biāo)記兔抗人IgAMulti-class antibodies規(guī)格： 0.1ml

軟骨糖蛋白39抗體 Anti-CDR/YKL-40 0.1ml

Rabbit Anti-horse IgM/Cy7 Cy7標(biāo)記的兔抗馬IgM 0.1ml

FAM29A 英文名稱： FAM29蛋白抗體 0.2ml

Rhesus antibody Rh PLAGL1/ZAC1 多型性腺瘤基因樣蛋白1抗體 規(guī)格 0.2ml

Rabbit Anti-human IgA/HRP 辣根過氧化物酶標(biāo)記兔抗人IgAMulti-class antibodies規(guī)格： 0.1ml

Sca1 英文名稱： T淋巴細(xì)胞激活蛋白抗體 0.2ml

LPA1 英文名稱： 溶血磷脂酸受體蛋白1抗體 0.1ml

一種新的絲氨酸/蘇氨酸激酶家族成員的基因WNK4抗體 Anti-WNK4 0.2ml

Anti-TSHR(NT)/FITC 熒光素標(biāo)記促甲狀腺素受體抗體(N端)IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh phospho-IKK beta(Ser733) 磷酸化KB抑制蛋白激酶β抗體 規(guī)格 0.1ml

TRA16 (Testicular orphan nuclear receptor-4 (TR4)-associated protein) 激素受體相關(guān)蛋白Multi-class antibodies規(guī)格： 0.5mg

HBE, 正常人上皮細(xì)胞系

DQSHS1 迪慶綿羊皮膚成纖維樣細(xì)胞

OMG Others Human 人 OMG / OMGP (aa 1-416) 人細(xì)胞裂解液 (陽性對(duì)照)

CM-M062小鼠腎小管上皮細(xì)胞完全培養(yǎng)基100mL

成纖維細(xì)胞培養(yǎng)基FM-acf-prf

EB病毒轉(zhuǎn)化的人B淋巴細(xì)胞;KMY0903 新生兒細(xì)胞完全培養(yǎng)基 100mL

CL-0068COS-1(非洲綠猴SV40轉(zhuǎn)化的腎細(xì)胞)5×106cells/瓶×2

PTPRC Others Mouse 小鼠 CD45 / PTPRC 人細(xì)胞裂解液 (陽性對(duì)照)

CCCSCK

8910PM細(xì)胞，人細(xì)胞 小鼠細(xì)胞,EMT6細(xì)胞 腎小管上皮細(xì)胞Many types of cells包裝：5 × 105次方(1ml)

Dami(人巨核細(xì)胞細(xì)胞) 5×106cells/瓶×2

HTEpC Pellet 人類氣管上皮細(xì)胞團(tuán)塊 > 1 mio.cells 人骨骼肌細(xì)胞cDNAHSkMC cDNA

Ras樣蛋白A抗體說明書 HBE, 正常人上皮細(xì)胞系

DQSHS1 迪慶綿羊皮膚成纖維樣細(xì)胞

OMG Others Human 人 OMG / OMGP (aa 1-416) 人細(xì)胞裂解液 (陽性對(duì)照)

CM-M062小鼠腎小管上皮細(xì)胞完全培養(yǎng)基100mL

成纖維細(xì)胞培養(yǎng)基FM-acf-prf

EB病毒轉(zhuǎn)化的人B淋巴細(xì)胞;KMY0903 新生兒細(xì)胞完全培養(yǎng)基 100mL