抗体的生物素化标记实验要点：

1. 抑制Hh信号通路蛋白SUFU抗体品牌 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-SUFU/Suppressor of Fused 

中文名称   抑制Hh信号通路蛋白SUFU抗体品牌 

别    名  SU FU; SU(F)U; Su(fu); SUFU; SUFU_HUMAN; SUFUH; SUFUXL; Suppressor of fused homolog (Drosophila); Suppressor of fused homolog; OTTHUMP00000020374; OTTHUMP00000020377; OTTHUMP00000020379; PRO1280.

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, Chicken, Dog, Pig, Horse 

产品类型   一抗  

研究领域     转录调节因子 表观遗传学

蛋白分子量  predicted molecular weight: 53, 106kDa

性    状   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human SUFU(433-485 aa) 

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

抑制Hh信号通路蛋白SUFU抗体品牌 产品应用    ELISA=1:500-1000 IP=1:20-100 IHC-P=1:100-500 IHC-F=1:100-500 ICC=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 SUFU is a key negative regulator in the vertebrate Hedgehog signaling pathway. SUFU interacts with genes encoding proteins in this signal transduction pathway. In Drosophila, Intracellular transduction of the Hedgehog pathway involves the release of a large complex containing SUFU. SUFU inhibits the activity of the transcription factor Gli1 and interacts with Gli2, Gli3 and the serine/threonine kinase Fused. SUFU is widely expressed in adult and embryonic tissues with higher expression in tissues patterned by hegdehog signaling. The SUFU gene locus maps to a region that is deleted in glioblastomas, prostate cancer, malignant melanoma and endometrial cancer.

Function : Negative regulator in the hedgehog signaling pathway. Down-regulates GLI1-mediated transactivation of target genes. Part of a corepressor complex that acts on DNA-bound GLI1. May also act by linking GLI1 to BTRC and thereby targeting GLI1 to degradation by the proteasome. Sequesters GLI1, GLI2 and GLI3 in the cytoplasm, this effect is overcome by binding of STK36 to both SUFU and a GLI protein. Negative regulator of beta-catenin signaling. Regulates the formation of either the repressor form (GLI3R) or the activator form (GLI3A) of the full length form of GLI3 (GLI3FL). GLI3FL is complexed with SUFU in the cytoplasm and is maintained in a neutral state. Without the Hh signal, the SUFU-GLI3 complex is recruited to cilia, leading to the efficient processing of GLI3FL into GLI3R. When Hh signaling is initiated, SUFU dissociates from GLI3FL and the latter translocates to the nucleus, where it is phosphorylated, destabilized, and converted to a transcriptional activator (GLI3A).

Subunit : May form homodimers. Part of a DNA-bound corepressor complex containing SAP18, GLI1 and SIN3. Part of a complex containing CTNNB1. Binds BTRC, GLI2, GLI3, SAP18 and STK36. Binds both free and DNA-bound GLI1. Interacts with KIF7. Interacts with GLI3FL and this interaction regulates the formation of either repressor or activator forms of GLI3. Its association with GLI3FL is regulated by Hh signaling and dissociation of the SUFU-GLI3 interaction requires the presence of the ciliary motor KIF3A. Interacts with ULK3; inactivating the protein kinase activity of ULK3.

Subcellular Location : Cytoplasm. Nucleus.

Tissue Specificity : Ubiquitous in adult tissues. Detected in osteoblasts of the perichondrium in the developing limb of 12-week old embryos. Isoform 1 is detected in fetal brain, lung, kidney and testis. Isoform 2 is detected in fetal testis, and at much lower levels in fetal brain, lung and kidney.

DISEASE : Defects in SUFU are a cause of medulloblastoma (MDB) [MIM:155255]. MDB is a malignant, invasive embryonal tumor of the cerebellum with a preferential manifestation in children. Defects in SUFU play a role in predisposition to desmoplastic MDB. These tumors make up about 20 to 30% of medulloblastomas, have a more nodular architecture than 'classical' medulloblastoma, and may have a better prognosis.

Similarity : Belongs to the SUFU family.

抗体的鉴定：

1）抑制Hh信号通路蛋白SUFU抗体品牌 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。

哥伦比亚琼脂培养基2406g灭菌后冷至45℃左右时加入5%脱纤维羊血，用于空肠弯曲菌的培养。(CP、GB、EP、USP)

2561-prf TEpiCM-prf 扁桃体上皮细胞培养基 500ml incubation media 2561-prf TEpiCM-prf 扁桃体上皮细胞培养基 500ml

改良马丁培养基 300ml/袋*10 用于药品及生物制品霉菌无菌检验

Pfizer肠球菌选择性琼脂(PSE琼脂)250g用于粪链球菌的选择性分离和计数(SN标准)

GCAgar

哥伦比亚琼脂培养基2406g用于营养要求较高的细菌的培养及溶血试验，还用于药品中梭菌的检测。(《中华人民共和国药典》201...

TYGPN Medium 培养基 250g incubation media TYGPN Medium 培养基 250g

玉蕈、真姬菇 支/瓶

肠道菌增菌肉汤(EE)250g用于肠道菌的增菌培养(FDA、SN标准)

GVPC琼脂基础添加剂 5支/套 每套添加于100mlGVPC琼脂中

大豆酪蛋白消化物琼脂培养基250g用于药品中需氧菌总数测定

mEI琼脂(肠球菌显色培养基) 1000mL/瓶 incubation media mEI琼脂(肠球菌显色培养基) 1000mL/瓶

哥伦比亚琼脂培养基 250g 用于营养要求较高的细菌的培养及溶血试验，还用于药品中梭菌的检测。(《中华人民共和国药典》201...

琼脂培养基JAgarmediumJ(Deoxycholatecitrateagar)用于沙门氏菌选择性分离培养

M17Agar

CL-0236U-2 OS(人细胞)5×106cells/瓶×2

IL1R1 Others Human 人 IL1R1 / CD121a 人细胞裂解液 (阳性对照)

人肺动脉成纤维细胞cDNAHPAF cDNA

HCC38细胞，人导管癌细胞 昆虫细胞系,SF-9细胞 肝动脉内皮细胞Many types of cells包装：5 × 105次方(1ml)

293E(人胚肾细胞(EBNA1基因修饰)) 5×106cells/瓶×2

HBMEC Pellet 正常人脑部微内皮细胞团块 > 1 mio.cells EpiFectagen? 上皮细胞转染试剂盒

抑制Hh信号通路蛋白SUFU抗体品牌CL-0234TM3(小鼠间质细胞)5×106cells/瓶×2

KIT Others Human 人 KIT / c-KIT / CD117 人细胞裂解液 (阳性对照)

人肺动脉内皮细胞cDNAHPAEC cDNA

小鼠髋动脉内皮细胞;PIEC 细胞，RBL-2H3细胞 ME细胞，C57小鼠瘤株

Sars结构蛋白表达株;293 001B

HA Others H1N1 甲型 H1N1 (A/Puerto Rico/8/1934) 血凝素 (Hemagglutinin / HA) 人细胞裂解液 (阳性对照)