抗体的生物素化标记实验要点：

1. 磷酸化核仁磷酸蛋白抗体规格 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*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-NPM (Thr199) 

中文名称   磷酸化核仁磷酸蛋白抗体规格 

别    名  Nucleophosmin (phospho T199); Nucleophosmin (phospho Thr199); p-Nucleophosmin (Thr199); B23; B23; MGC104254; NMP1; NMP1; NO38; NPM 1; NPM; NPM_HUMAN; NPM1; Nucleolar Phosphoprotein B23; Nucleolar Phosphoprotein B23; Nucleolar protein NO38; Nucleophosmin (nucleolar phosphoprotein B23 numatrin); Nucleophosmin; Nucleophosmin/B23.2; Nucleophosmin/nucleoplasmin family member 1; Nucleoplasmin Family Member 1; Nucleoplasmin Family Member 1; Numatrin; Numatrin; OTTHUMP00000161024; OTTHUMP00000161025; OTTHUMP00000223397; OTTHUMP00000223398; TRK fused gene.

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

产品类型   一抗 磷酸化抗体  

研究领域     细胞生物 免疫学 神经生物学 信号转导 细胞周期蛋白 转录调节因子

蛋白分子量  predicted molecular weight: 38kDa

性    状   Lyophilized or Liquid

免 疫 原 KLH conjugated Synthesised phosphopeptide derived from human NPM around the phosphorylation site of Thr199 

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

磷酸化核仁磷酸蛋白抗体规格 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 This gene encodes a phosphoprotein which moves between the nucleus and the cytoplasm. The gene product is thought to be involved in several processes including regulation of the ARF/p53 pathway. A number of genes are fusion partners have been characterized, in particular the anaplastic lymphoma kinase gene on chromosome 2. Mutations in this gene are associated with acute myeloid leukemia. More than a dozen pseudogenes of this gene have been identified. Alternative splicing results in multiple transcript variants.[provided by RefSeq, Nov 2009].

Function : Involved in diverse cellular processes such as ribosome biogenesis, centrosome duplication, protein chaperoning, histone assembly, cell proliferation, and regulation of tumor suppressors p53/TP53 and ARF. Binds ribosome presumably to drive ribosome nuclear export. Associated with nucleolar ribonucleoprotein structures and bind single-stranded nucleic acids. Acts as a chaperonin for the core histones H3, H2B and H4. Stimulates APEX1 endonuclease activity on apurinic/apyrimidinic (AP) double-stranded DNA but inhibits APEX1 endonuclease activity on AP single-stranded RNA. May exert a control of APEX1 endonuclease activity within nucleoli devoted to repair AP on rDNA and the removal of oxidized rRNA molecules.Subunit : Nucleus, nucleolus. Nucleus, nucleoplasm. Note=Generally nucleolar, but is translocated to the nucleoplasm in case of serum starvation or treatment with anticancer drugs. Has been found in the cytoplasm in patients with primary acute myelogenous leukemia (AML), but not with secondary AML. Can shuttle between cytoplasm and nucleus. Co-localizes with the methylated form of RPS10 in the granular component (GC) region of the nucleolus. Colocalized with nucleolin and APEX1 in nucleoli.Post-translational modifications : Acetylated at C-terminal lysine residues, thereby increasing affinity to histones.ADP-ribosylated.Phosphorylated at Ser-4 by PLK1. Phosphorylated by CDK2 at Ser-125 and Thr-199. Phosphorylation at Thr-199 may trigger initiation of centrosome duplication. Phosphorylated by CDK1 at Thr-199, Thr-219, Thr-234 and Thr-237 during cell mitosis. When these four sites are phosphorated, RNA-binding activity seem to be abolished. May be phosphorylated at Ser-70 by NEK2.Sumoylated by ARF.DISEASE : Note=A chromosomal aberration involving NPM1 is found in a form of non-Hodgkin lymphoma. Translocation t(2;5)(p23;q35) with ALK. The resulting chimeric NPM1-ALK protein homodimerize and the kinase becomes constitutively activated.Note=A chromosomal aberration involving NPM1 is found in a form of acute promyelocytic leukemia. Translocation t(5;17)(q32;q11) with RARA.Note=A chromosomal aberration involving NPM1 is a cause of myelodysplastic syndrome (MDS). Translocation t(3;5)(q25.1;q34) with MLF1.Note=Defects in NPM1 are associated with acute myelogenous leukemia (AML). Mutations in exon 12 affecting the C-terminus of the protein are associated with an aberrant cytoplasmic location.Similarity : Belongs to the nucleoplasmin family.Database links : UniProtKB/Swiss-Prot: P06748.2核仁磷酸蛋白（nucleophosmin）是雌激素调控的一种核仁蛋白， Nucleophosmin(又称为NPM、B23、Numatrin、NO38)位于核仁的颗粒区,与其他一些核蛋白如C23、P120等可相互作用。NPM可以在细胞核与细胞浆之间穿梭,其定位受细胞周期及一些细胞*物的影响。NPM基因在多种人类中过表达,因此被公认为标志物和癌基因.后来研究发现NPM也具有,它的缺失、突变甚至重排与多种的发生密切相关.

抗体的鉴定：

1）磷酸化核仁磷酸蛋白抗体规格 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。

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Rhesus antibody Rh CCR-2B 细胞表面趋化因子受体2B抗体 规格 0.1ml

Inhibin beta A peptide 抑制素beta A抗原 0.5mg

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ANG-4(Mouse Angiopoietin 4) ELISA Kit 小鼠生成素4Multi-class antibodies规格： 48T

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Rhesus antibody Rh Endothelial lipase 内皮脂肪酶抗体 规格 0.2ml

PYD ELISA Kit 大鼠酚 96T

MS4A15 英文名称： MS4A15蛋白抗体 0.1ml

phospho-ATF2 (Ser44) 英文名称： 磷酸化活化复制因子2抗体 0.1ml

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Anti-Melamine/Biotin 生物素标记兔抗三聚氰胺抗体IgGMulti-class antibodies规格： 0.2ml

Anti-obestatin/Ghrelin/GHRP /FITC 荧光素标记抑制素抗体IgGMulti-class antibodies规格： 0.2ml

Rhesus antibody Rh BMI-1/PCGF4 组蛋白相关Bmi1抗体 规格 0.2ml

Gastrin 胃泌素(抗原) 0.5mg

HLA-DR 英文名称： HLA-DR抗体 0.1ml

Rhesus antibody Rh R-cadherin/Cadherin 4 R-钙粘附分子抗体 规格 0.1ml

Anti-obestatin/Ghrelin/GHRP /FITC 荧光素标记抑制素抗体IgGMulti-class antibodies规格： 0.2ml

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