抗体的生物素化标记实验要点：

1. 磷酸化指状蛋白RET抗体规格 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-Phospho-Ret (Tyr1062) 

中文名称   磷酸化指状蛋白RET抗体规格 

别    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.

浓    度   1mg/1ml

规 格  0.1ml/100μg

抗体来源   Rabbit

克隆类型   polyclonal

交叉反应   Human, Mouse, Rat, Dog, Cow, Rabbit 

产品类型   一抗 磷酸化抗体  

研究领域     细胞生物 免疫学 信号转导 生长因子和激素 激酶和磷酸酶

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

性    状   Lyophilized or Liquid

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

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

磷酸化指状蛋白RET抗体规格 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 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指状蛋白属于一个较大的B-盒环指蛋白家族，RET与酪氨酸激酶融合后就变为癌基因.

抗体的鉴定：

1）磷酸化指状蛋白RET抗体规格 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。

HLA-E (Human Leukocyte Anti-gen-E) 人类白细胞抗原EMulti-class antibodies规格： 0.5mg

Anti-AACT-Alpha1 α-1抗胰糜蛋白酶抗体Multi-class antibodies规格： 0.1ml

Rhesus antibody Rh phospho-c-Jun(Tyr170) 磷酸化原癌基因c-Jun抗体 规格 0.1ml

羊抗人IgM血清 0.5ml 国产

WWOX 英文名称： 包含氧化还原酶的WW域抗体 0.1ml

DcR3 英文名称： 诱捕受体3抗体 0.1ml

Anti-AACT-Alpha1 α-1抗胰糜蛋白酶抗体Multi-class antibodies规格： 0.1ml

Anti-TRF/Gold 金离子标记抗转铁蛋白抗体IgGMulti-class antibodies规格： 0.5ml(15nm)

rhEGF(rh-Epidermal growth factor) 重组人表皮生长因子Multi-class antibodies规格： 100ug

β-抑制蛋白2抗体 Anti-β-arrestin 2 0.1ml

ZNF93 英文名称： 锌指蛋白93抗体 0.2ml

DNAH9 英文名称： 轴丝动力蛋白重链9抗体 0.2ml

Rhesus antibody Rh phospho-eIF4EBP1(Thr36) 磷酸化4E结合蛋白1抗体 规格 0.1ml

rhEGF(rh-Epidermal growth factor) 重组人表皮生长因子Multi-class antibodies规格： 100ug

5 lipoxygenase 英文名称： 5-脂氧合酶抗体 0.1ml

phospho-ECT2 (Ser370) 英文名称： 磷酸化上皮细胞癌转化蛋白2抗体 0.1ml

反应性溶血膜抑制蛋白抗体 Anti-CD59/potectin 0.1ml

Anti-Phospho-FLT3 (Tyr591) /FITC 荧光素标记兔抗人、大、小鼠磷酸化FMS样酪氨酸激酶3IgGMulti-class antibodies规格： 0.2ml

Rhesus antibody Rh phospho-GFAP (Ser8) 磷酸化胶质纤维酸性蛋白抗体 规格 0.1ml

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

AIMP1 Protein Human 重组人 AIMP1 / EMAP2 / SCYE1 蛋白 (His 标签)

MS751 人子宫颈表皮癌细胞

CD93 Others Human 人 CD93 / C1QR1 人细胞裂解液 (阳性对照)

NCI-H661人大细胞细胞 NCI-H661 large cell lung cancer cell RPMI-1640(GIBCO)+10%FBS

人慢性髓系细胞;K562

人胚肺二倍体细胞;2BS 人成纤维细胞完全培养基 100mL

CL-0256AM(人腺样囊性癌细胞(高转移))5×106cells/瓶×2

SCN3B Others Human 人 SCN3B 人细胞裂解液 (阳性对照)

小鼠*上皮细胞完全培养基 100mL

B16小鼠细胞 B16 mouse melanoma cells RPMI-1640(GIBCO)+10%FBS

IL33 Protein Human 重组人 IL33 / Ierleukin-33 / NF-HEV 蛋白

PC-3M-2B4(人低转移细胞株) 5×106cells/瓶×2 CHO/dhFr-(中国仓鼠二氢叶酸还原酶缺陷细胞)

磷酸化指状蛋白RET抗体规格 AIMP1 Protein Human 重组人 AIMP1 / EMAP2 / SCYE1 蛋白 (His 标签)

MS751 人子宫颈表皮癌细胞

CD93 Others Human 人 CD93 / C1QR1 人细胞裂解液 (阳性对照)

NCI-H661人大细胞细胞 NCI-H661 large cell lung cancer cell RPMI-1640(GIBCO)+10%FBS

人慢性髓系细胞;K562

人胚肺二倍体细胞;2BS 人成纤维细胞完全培养基 100mL