抗體的生物素化標(biāo)記實(shí)驗(yàn)要點(diǎn)：

1. 磷酸化指狀蛋白R(shí)ET抗體規(guī)格 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(lái)篩選最適條件；

3.用NHSB量過(guò)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(gòu)信息：
英文名稱  Anti-Phospho-Ret (Tyr1062) 

中文名稱   磷酸化指狀蛋白R(shí)ET抗體規(guī)格 

別    名  C ret; Cadherin family member 12; Cadherin related family member 16; CDHF 12; CDHF12; CDHR16; ELKS Fusion gene; HSCR 1; HSCR1; Hydroxyaryl protein kinase; MEN2A; MEN2B; MTC 1; MTC1; Multiple endocrine neoplasia and medullary thyroid carcinoma 1; Oncogene RET; Proto oncogene tyrosine protein kinase receptor ret; Proto-oncogene c-Ret; Proto-oncogene tyrosine-protein kinase receptor ret; PTC; RET; RET ELE1; Ret Proto oncogene; RET transforming sequence; RET_HUMAN; RET51; RET9; tyrosine-protein kinase receptor ret.

濃    度   1mg/1ml

規(guī) 格  0.1ml/100μg

抗體來(lái)源   Rabbit

克隆類型   polyclonal

交叉反應(yīng)   Human, Mouse, Rat, Dog, Cow, Rabbit 

產(chǎn)品類型   一抗 磷酸化抗體  

研究領(lǐng)域     細(xì)胞生物 免疫學(xué) 信號(hào)轉(zhuǎn)導(dǎo) 生長(zhǎng)因子和激素 激酶和磷酸酶

蛋白分子量  predicted molecular weight: 34/76/122kDa

性    狀   Lyophilized or Liquid

免 疫 原 KLH conjugated Synthesised phosphopeptide derived from human Ret around the phosphorylation site of Tyr1062 

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

磷酸化指狀蛋白R(shí)ET抗體規(guī)格 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 The CD4 antigen is highly expressed on human T helper cells and thymocytes, and at lower levels on monocytes and dendritic cells. This integral membrane glycoprotein of approximately 58 kDa contains four external domains (D1 to D4) that show homology to members of the immunoglobulin (Ig) superfamily. It is responsible for the recognition of the MHC class II antigen. The CD4 antibody recognizes most thymocytes and about 65% of peripheral blood T cells.

Function : Receptor tyrosine-protein kinase involved in numerous cellular mechanisms including cell proliferation, neuronal navigation, cell migration, and cell differentiation upon binding with glial cell derived neurotrophic factor family ligands. Phosphorylates PTK2/FAK1. Regulates both cell death/survival balance and positional information. Required for the molecular mechanisms orchestration during intestine organogenesis; involved in the development of enteric nervous system and renal organogenesis during embryonic life, and promotes the formation of Peyer's patch-like structures, a major component of the gut-associated lymphoid tissue. Modulates cell adhesion via its cleavage by caspase in sympathetic neurons and mediates cell migration in an integrin (e.g. ITGB1 and ITGB3)-dependent manner. Involved in the development of the neural crest. Active in the absence of ligand, triggering apoptosis through a mechanism that requires receptor intracellular caspase cleavage. Act as a dependence receptor; in the presence of the ligand GDNF in somatotrophs (within pituitary), promotes survival and down regulates growth hormone (GH) production, but triggers apoptosis in absence of GDNF. Regulates nociceptor survival and size. Triggers the differentiation of rapidly adapting (RA) mechanoreceptors. Mediator of several diseases such as neuroendocrine cancers; these diseases are characterized by aberrant integrins-regulated cell migration.

Subunit : Phosphorylated form interacts with the PBT domain of DOK2, DOK4 and DOK5. The phosphorylated form interacts with PLCG1 and GRB7 (By similarity). Interacts (not phosphorylated) with CC PTK2/FAK1 (via FERM domain). Extracellular cell-membrane anchored RET cadherin fragments form complex in neurons with reduced trophic status, preferentially at the contact sites between somas. Interacts with AIP in the pituitary gland; this interaction prevents the formation of the AIP-survivin complex. Binds to ARTN.

Subcellular Location : Cell membrane; Single-pass type I membrane protein. Endosome membrane; Single-pass type I membrane protein.

Post-translational modifications : Autophosphorylated on C-terminal tyrosine residues upon ligand stimulation. Dephosphorylated by PTPRJ on Tyr-905, Tyr-1015 and Tyr-1062.

Proteolytically cleaved by caspase-3. The soluble RET kinase fragment is able to induce cell death. The extracellular cell-membrane anchored RET cadherin fragment accelerates cell adhesion in sympathetic neurons.

DISEASE : Defects in RET may be a cause of colorectal cancer (CRC) [MIM:114500].

Defects in RET are a cause of Hirschsprung disease type 1 (HSCR1) [MIM:142623]. HSCR1 is a disorder of neural crest development characterized by the absence of intramural ganglion cells in the myenteric and submucosal plexuses of the gastrointestinal tract, often resulting in intestinal obstruction. Total colonic aganglionosis and total intestinal Hirschsprung disease also occur. Occasionally, MEN2A or FMTC occur in association with HSCR1.

Defects in RET are the cause of medullary thyroid carcinoma (MTC) [MIM:155240]. MTC is a rare tumor derived from the C cells of the thyroid. Three hereditary forms are known, that are transmitted in an autosomal dominant fashion: (a) multiple neoplasia type 2A (MEN2A), (b) multiple neoplasia type IIB (MEN2B) and (c) familial MTC (FMTC), which occurs in 25-30% of MTC cases and where MTC is the only clinical manifestation.

Defects in RET are the cause of multiple neoplasia type 2B (MEN2B) [MIM:162300]. MEN2B is an uncommon inherited cancer syndrome characterized by predisposition to MTC and phaeochromocytoma which is associated with marfanoid habitus, mucosal neuromas, skeletal and ophtalmic abnormalities, and ganglioneuromas of the intestine tract. Then the disease progresses rapidly with the development of metastatic MTC and a pheochromocytome in 50% of cases.

Defects in RET are a cause of susceptibility to pheochromocytoma (PCC) [MIM:171300]. A catecholamine-producing tumor of chromaffin tissue of the adrenal medulla or sympathetic paraganglia. The cardinal symptom, reflecting the increased secretion of epinephrine and norepinephrine, is hypertension, which may be persistent or intermittent.

Defects in RET are the cause of multiple neoplasia type 2A (MEN2A) [MIM:171400]; also known as multiple neoplasia type 2 (MEN2). MEN2A is the most frequent form of medullary thyroid cancer (MTC). It is an inherited cancer syndrome characterized by MTC, phaeochromocytoma and/or hyperparathyroidism.

Defects in RET are a cause of thyroid papillary carcinoma (TPC) [MIM:188550]. TPC is a common tumor of the thyroid that typically arises as an irregular, solid or cystic mass from otherwise normal thyroid tissue. Papillary carcinomas are malignant neoplasm characterized by the formation of numerous, irregular, finger-like projections of fibrous stroma that is covered with a surface layer of neoplastic epithelial cells. Note=Chromosomal aberrations involving RET are found in thyroid papillary carcinomas. Inversion inv(10)(q11.2;q21) generates the RET/CCDC6 (PTC1) oncogene; inversion inv(10)(q11.2;q11.2) generates the RET/NCOA4 (PTC3) oncogene; translocation t(10;14)(q11;q32) with GOLGA5 generates the RET/GOLGA5 (PTC5) oncogene; translocation t(8;10)(p21.3;q11.2) with PCM1 generates the PCM1/RET fusion; translocation t(6;10)(p21.3;q11.2) with RFP generates the Delta RFP/RET oncogene; translocation t(1;10)(p13;q11) with TRIM33 generates the TRIM33/RET (PTC7) oncogene; translocation t(7;10)(q32;q11) with TRIM24/TIF1 generates the TRIM24/RET (PTC6) oncogene. The PTC5 oncogene has been found in 2 cases of PACT in children exposed to radioactive fallout after Chernobyl. A chromosomal aberration involving TRIM27/RFP is found in thyroid papillary carcinomas. Translocation t(6;10)(p21.3;q11.2) with RET. The translocation generates TRIM27/RET and delta TRIM27/RET oncogenes.

Defects in RET are a cause of renal adysplasia (RADYS) [MIM:191830]; also known as renal agenesis or renal aplasia. Renal agenesis refers to the absence of one (unilateral) or both (bilateral) kidneys at birth. Bilateral renal agenesis belongs to a group of perinatally lethal renal diseases, including severe bilateral renal dysplasia, unilateral renal agenesis with contralateral dysplasia and severe obstructive uropathy.

Defects in RET are a cause of congenital central hypoventilation syndrome (CCHS) [MIM:209880]; also known as congenital failure of autonomic control or Ondine curse. CCHS is a rare disorder characterized by abnormal control of respiration in the absence of neuromuscular or lung disease, or an identifiable brain stem lesion. A deficiency in autonomic control of respiration results in inadequate or negligible ventilatory and arousal responses to hypercapnia and hypoxemia.

Similarity : Belongs to the protein kinase superfamily. Tyr protein kinase family.

Contains 1 cadherin domain.

Contains 1 protein kinase domain.

Database links : UniProtKB/Swiss-Prot: P07949.3

RET指狀蛋白屬于一個(gè)較大的B-盒環(huán)指蛋白家族，RET與酪氨酸激酶融合后就變?yōu)榘┗?

抗體的鑒定：

1）磷酸化指狀蛋白R(shí)ET抗體規(guī)格 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(lái)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(lái)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(wú)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(kù)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。

HLA-E (Human Leukocyte Anti-gen-E) 人類白細(xì)胞抗原EMulti-class antibodies規(guī)格： 0.5mg

Anti-AACT-Alpha1 α-1抗胰糜蛋白酶抗體Multi-class antibodies規(guī)格： 0.1ml

Rhesus antibody Rh phospho-c-Jun(Tyr170) 磷酸化原癌基因c-Jun抗體 規(guī)格 0.1ml

羊抗人IgM血清 0.5ml 國(guó)產(chǎn)

WWOX 英文名稱： 包含氧化還原酶的WW域抗體 0.1ml

DcR3 英文名稱： 誘捕受體3抗體 0.1ml

Anti-AACT-Alpha1 α-1抗胰糜蛋白酶抗體Multi-class antibodies規(guī)格： 0.1ml

Anti-TRF/Gold 金離子標(biāo)記抗轉(zhuǎn)鐵蛋白抗體IgGMulti-class antibodies規(guī)格： 0.5ml(15nm)

rhEGF(rh-Epidermal growth factor) 重組人表皮生長(zhǎng)因子Multi-class antibodies規(guī)格： 100ug

β-抑制蛋白2抗體 Anti-β-arrestin 2 0.1ml

ZNF93 英文名稱： 鋅指蛋白93抗體 0.2ml

DNAH9 英文名稱： 軸絲動(dòng)力蛋白重鏈9抗體 0.2ml

Rhesus antibody Rh phospho-eIF4EBP1(Thr36) 磷酸化4E結(jié)合蛋白1抗體 規(guī)格 0.1ml

rhEGF(rh-Epidermal growth factor) 重組人表皮生長(zhǎng)因子Multi-class antibodies規(guī)格： 100ug

5 lipoxygenase 英文名稱： 5-脂氧合酶抗體 0.1ml

phospho-ECT2 (Ser370) 英文名稱： 磷酸化上皮細(xì)胞癌轉(zhuǎn)化蛋白2抗體 0.1ml

反應(yīng)性溶血膜抑制蛋白抗體 Anti-CD59/potectin 0.1ml

Anti-Phospho-FLT3 (Tyr591) /FITC 熒光素標(biāo)記兔抗人、大、小鼠磷酸化FMS樣酪氨酸激酶3IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh phospho-GFAP (Ser8) 磷酸化膠質(zhì)纖維酸性蛋白抗體 規(guī)格 0.1ml

CEA(Carcinoembryonic Antigen )peptide 人癌胚抗原抗原Multi-class antibodies規(guī)格： 0.5mg

AIMP1 Protein Human 重組人 AIMP1 / EMAP2 / SCYE1 蛋白 (His 標(biāo)簽)

MS751 人子宮頸表皮癌細(xì)胞

CD93 Others Human 人 CD93 / C1QR1 人細(xì)胞裂解液 (陽(yáng)性對(duì)照)

NCI-H661人大細(xì)胞細(xì)胞 NCI-H661 large cell lung cancer cell RPMI-1640(GIBCO)+10%FBS

人慢性髓系細(xì)胞;K562

人胚肺二倍體細(xì)胞;2BS 人成纖維細(xì)胞完全培養(yǎng)基 100mL

CL-0256AM(人腺樣囊性癌細(xì)胞(高轉(zhuǎn)移))5×106cells/瓶×2

SCN3B Others Human 人 SCN3B 人細(xì)胞裂解液 (陽(yáng)性對(duì)照)

小鼠*上皮細(xì)胞完全培養(yǎng)基 100mL

B16小鼠細(xì)胞 B16 mouse melanoma cells RPMI-1640(GIBCO)+10%FBS

IL33 Protein Human 重組人 IL33 / Ierleukin-33 / NF-HEV 蛋白

PC-3M-2B4(人低轉(zhuǎn)移細(xì)胞株) 5×106cells/瓶×2 CHO/dhFr-(中國(guó)倉(cāng)鼠二氫葉酸還原酶缺陷細(xì)胞)

磷酸化指狀蛋白R(shí)ET抗體規(guī)格 AIMP1 Protein Human 重組人 AIMP1 / EMAP2 / SCYE1 蛋白 (His 標(biāo)簽)

MS751 人子宮頸表皮癌細(xì)胞

CD93 Others Human 人 CD93 / C1QR1 人細(xì)胞裂解液 (陽(yáng)性對(duì)照)

NCI-H661人大細(xì)胞細(xì)胞 NCI-H661 large cell lung cancer cell RPMI-1640(GIBCO)+10%FBS

人慢性髓系細(xì)胞;K562

人胚肺二倍體細(xì)胞;2BS 人成纖維細(xì)胞完全培養(yǎng)基 100mL