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抗體的生物素化標(biāo)記實(shí)驗(yàn)要點(diǎn)：

1. 磷酸化核仁磷酸蛋白抗體規(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-NPM (Thr199) 

中文名稱   磷酸化核仁磷酸蛋白抗體規(guī)格 

別    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.

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

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) 神經(jīng)生物學(xué) 信號(hào)轉(zhuǎn)導(dǎo) 細(xì)胞周期蛋白 轉(zhuǎn)錄調(diào)節(jié)因子

蛋白分子量  predicted molecular weight: 38kDa

性    狀   Lyophilized or Liquid

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

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

磷酸化核仁磷酸蛋白抗體規(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 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）是雌激素調(diào)控的一種核仁蛋白， Nucleophosmin(又稱為NPM、B23、Numatrin、NO38)位于核仁的顆粒區(qū),與其他一些核蛋白如C23、P120等可相互作用。NPM可以在細(xì)胞核與細(xì)胞漿之間穿梭,其定位受細(xì)胞周期及一些細(xì)胞*物的影響。NPM基因在多種人類中過(guò)表達(dá),因此被公認(rèn)為標(biāo)志物和癌基因.后來(lái)研究發(fā)現(xiàn)NPM也具有,它的缺失、突變甚至重排與多種的發(fā)生密切相關(guān).

抗體的鑒定：

1）磷酸化核仁磷酸蛋白抗體規(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。抗體的特異性高,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。

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Anti-Insulin(1-51 protein)/FITC 熒光素標(biāo)記1-51蛋白抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh CCR-2B 細(xì)胞表面趨化因子受體2B抗體 規(guī)格 0.1ml

Inhibin beta A peptide 抑制素beta A抗原 0.5mg

Phospho-IRAK1 (Thr387) 英文名稱： 磷酸化白介素-1受體相關(guān)激酶1抗體 0.1ml

Rhesus antibody Rh SPINK1/SPINK3 相關(guān)胰蛋白酶抑制劑1抗體 規(guī)格 0.2ml

Anti-Insulin(1-51 protein)/FITC 熒光素標(biāo)記1-51蛋白抗體IgGMulti-class antibodies規(guī)格： 0.2ml

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

Anti-Calpain 1/FITC 熒光素標(biāo)記鈣蛋白酶抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh Endothelial lipase 內(nèi)皮脂肪酶抗體 規(guī)格 0.2ml

PYD ELISA Kit 大鼠酚 96T

MS4A15 英文名稱： MS4A15蛋白抗體 0.1ml

phospho-ATF2 (Ser44) 英文名稱： 磷酸化活化復(fù)制因子2抗體 0.1ml

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Anti-Melamine/Biotin 生物素標(biāo)記兔抗三聚氰胺抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Anti-obestatin/Ghrelin/GHRP /FITC 熒光素標(biāo)記抑制素抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh BMI-1/PCGF4 組蛋白相關(guān)Bmi1抗體 規(guī)格 0.2ml

Gastrin 胃泌素(抗原) 0.5mg

HLA-DR 英文名稱： HLA-DR抗體 0.1ml

Rhesus antibody Rh R-cadherin/Cadherin 4 R-鈣粘附分子抗體 規(guī)格 0.1ml

Anti-obestatin/Ghrelin/GHRP /FITC 熒光素標(biāo)記抑制素抗體IgGMulti-class antibodies規(guī)格： 0.2ml

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