抗体的生物素化标记实验要点：

1. 细胞周期检控Rad17蛋白抗体（复制因子C）费用 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-Rad17/RFC 

中文名称   细胞周期检控Rad17蛋白抗体（复制因子C）费用 

别    名  CCYC; Cell cycle checkpoint protein; Cell cycle checkpoint protein RAD17; HRAD 17; hRad17; R24L; Rad 17; Rad 24; RAD1 (S. pombe) homolog; RAD17; RAD17 homolog; Rad17 like protein; RAD17_HUMAN; RAD17Sp; Rad24; RF C activator 1 homolog; RF C/activator 1 homolog; RF-C/activator 1 homolog.

浓    度   1mg/1ml

规 格  0.1ml/100μg 0.2ml/200μg

抗体来源   Rabbit

克隆类型   polyclonal

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

产品类型   一抗  

研究领域    免疫学 染色质和核信号 表观遗传学

蛋白分子量  predicted molecular weight: 77kDa

性    状   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human Rad17 

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

细胞周期检控Rad17蛋白抗体（复制因子C）费用 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 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 The protein encoded by this gene is highly similar to the gene product of Schizosaccharomyces pombe rad17, a cell cycle checkpoint gene required for cell cycle arrest and DNA damage repair in response to DNA damage. This protein shares strong similarity with DNA replication factor C (RFC), and can form a complex with RFCs. This protein binds to chromatin prior to DNA damage and is phosphorylated by the checkpoint kinase ATR following damage. This protein recruits the RAD1-RAD9-HUS1 checkpoint protein complex onto chromatin after DNA damage, which may be required for its phosphorylation. The phosphorylation of this protein is required for the DNA-damage-induced cell cycle G2 arrest, and is thought to be a critical early event during checkpoint signaling in DNA-damaged cells. Multiple alternatively spliced transcript variants of this gene, which encode four distinct protein isoforms, have been reported. Two pseudogenes, located on chromosomes 7 and 13, have been identified. [provided by RefSeq, Jul 2013].

Function : Essential for sustained cell growth, maintenance of chromosomal stability, and ATR-dependent checkpoint activation upon DNA damage. Has a weak ATPase activity required for binding to chromatin. Participates in the recruitment of the RAD1-RAD9-HUS1 complex and C12orf32/RHINO onto chromatin, and in CHEK1 activation. May also serve as a sensor of DNA replication progression, and may be involved in homologous recombination.

Subunit : Part of a DNA-binding complex containing RFC2, RFC3, RFC4 and RFC5. Interacts with RAD1 and RAD9 within the RAD1-RAD9-HUS1 complex. Interacts with RAD9B, POLE, NHP2L1 and MCM7. DNA damage promotes interaction with ATR or ATM and disrupts interaction with the RAD1-RAD9-HUS1 complex.

Subcellular Location : Nucleus. Note=Phosphorylated form redistributes to discrete nuclear foci upon DNA damage.

Tissue Specificity : Overexpressed in various cancer cell lines and in colon carcinoma (at protein level). Isoform 2 and isoform 3 are the most abundant isoforms in non irradiated cells (at protein level). Ubiquitous at low levels. Highly expressed in testis, where it is expressed within the germinal epithelium of the seminiferous tubuli. Weakly expressed in seminomas (testicular tumors).

Post-translational modifications : Phosphorylated. Phosphorylation on Ser-646 and Ser-656 is cell cycle-regulated, enhanced by genotoxic stress, and required for activation of checkpoint signaling. Phosphorylation is mediated by ATR upon UV or replication arrest, whereas it may be mediated both by ATR and ATM upon ionizing radiation. Phosphorylation on both sites is required for interaction with RAD1 but dispensable for interaction with RFC3 or RFC4.

Similarity : Belongs to the rad17/RAD24 family.

Database links : UniProtKB/Swiss-Prot: O75943

抗体的鉴定：

1）细胞周期检控Rad17蛋白抗体（复制因子C）费用 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。

CD122/IL-2RB(Imterleukm-2 Receptor beta) CD122/白介素2受体β链抗原Multi-class antibodies规格： 0.5mg

Anti-beta-Amyloid(1-42) β淀粉样肽(1-42)抗体Multi-class antibodies规格： 0.1ml

Rhesus antibody Rh phospho-Bid (Ser65) 磷酸化BH3结构域凋亡诱导蛋白抗体 规格 0.1ml

ELISA试剂盒开发服务(10kit以内) 次 西唐

VWF 英文名称： 假性因子/性因子抗体 0.1ml

DPF2 英文名称： 双PHD-D4锌指蛋白2抗体 0.2ml

Anti-beta-Amyloid(1-42) β淀粉样肽(1-42)抗体Multi-class antibodies规格： 0.1ml

Anti-Phospho-PNK1 (Ser114/Thr118) /FITC 荧光素标记磷酸化多聚合苷酸激酶3磷酸化酶抗体IgGMulti-class antibodies规格： 0.2ml

Rabbit anti-mouse IgG1, kappa chain 兔抗小鼠IgG1k链 (亲和层析纯化)Multi-class antibodies规格： 1mg

葡萄糖调节蛋白94抗体 Anti-GRP94 0.1ml

Dog IgM/Cy5.5 Cy5.5标记的兔抗犬IgM抗体 0.1ml

FAM160B1 英文名称： FAM160B1蛋白抗体 0.2ml

Rhesus antibody Rh Phospho-S6 Ribosomal Protein (Ser235+Ser236) 磷酸化S6核糖体蛋白抗体 规格 0.1ml

Rabbit anti-mouse IgG1, kappa chain 兔抗小鼠IgG1k链 (亲和层析纯化)Multi-class antibodies规格： 1mg

SYNPR/Synaptoporin 英文名称： 突触囊泡蛋白抗体 0.2ml

EDG1 英文名称： 内皮细胞分化鞘脂G蛋白偶联受体1抗体 0.1ml

CREB结合蛋白抗体 Anti-CBP 0.1ml

Anti-ULBP1/N2DL1/FITC 荧光素标记兔抗人、大、小鼠UL16结合蛋白1蛋白抗体IgGMulti-class antibodies规格： 0.2ml

Rhesus antibody Rh Phospho-Histone H3 (Ser28) 磷酸化组蛋白H3抗体 规格 0.1ml

VEGF(Vascular endothelial growth factor) 内皮生长因子(多肽抗原)Multi-class antibodies规格： 0.5mg

RLF, 大鼠淋巴成纤维细胞

HEPG2.2.15 细胞HBV病毒株

FAP Others Human 人 FAP / Seprase 人细胞裂解液 (阳性对照)

CL-0290MDA-MB-468(人癌细胞)5×106cells/瓶×2

人间充质干细胞(骨髓)cDNAHMSC-bm cDNA

双位点HC-kit受体细胞株;DMF7 人子宫内膜上皮细胞完全培养基 100mL

CL-0154MDCK(犬肾细胞)5×106cells/瓶×2

IFNB1 Others Human 人 Ierferon beta / IFN-beta / IFNB CHO细胞裂解液 (阳性对照)

人肝窦内皮细胞裂解物HHSECL

小香猪皮肤细胞;SSC-S2 组织细胞细胞，U-937细胞 NUTU-19细胞，大鼠细胞株

SV40转染人成骨细胞;hFOB 1.19

HA Others H1N3 甲型 H1N3 (A/duck/NZL/160/1976) 血凝素HA1 (Hemagglutinin) 人细胞裂解液 (阳性对照)

细胞周期检控Rad17蛋白抗体（复制因子C）费用 RLF, 大鼠淋巴成纤维细胞

HEPG2.2.15 细胞HBV病毒株

FAP Others Human 人 FAP / Seprase 人细胞裂解液 (阳性对照)

CL-0290MDA-MB-468(人癌细胞)5×106cells/瓶×2

人间充质干细胞(骨髓)cDNAHMSC-bm cDNA

双位点HC-kit受体细胞株;DMF7 人子宫内膜上皮细胞完全培养基 100mL