抗體的生物素化標(biāo)記實(shí)驗(yàn)要點(diǎn)：

1. 抑制基因P53蛋白抗體（野生型P53）費(fèi)用 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)不同的分子比來篩選最適條件；

3.用NHSB量過量也是不利的,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)生競爭作用；

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)品訂購信息：
英文名稱  Anti-P53 protein(wt-p53) 

中文名稱   抑制基因P53蛋白抗體（野生型P53）費(fèi)用 

別    名  Widespread p53; Wtp53; Antigen NY-CO-13; Cellular tumor antigen p53; Cys 51 Stop; HGNC11998; LFS1; p53; p53 Cellular Tumor Antigen; p53 Tumor Suppressor; Phosphoprotein p53; TP53; Transformation related protein 53; TRP53; Tumor protein p53; Tumour Protein p53; P53_HUMAN; TP53.

濃    度   1mg/1ml

規(guī) 格  0.1ml/100μg 0.2ml/200μg

抗體來源   Rabbit

克隆類型   polyclonal

交叉反應(yīng)   Human, Mouse, Rat, Pig, Cow, Horse, Rabbit, Sheep 

產(chǎn)品類型   一抗  

研究領(lǐng)域     細(xì)胞生物 染色質(zhì)和核信號(hào) 細(xì)胞凋亡 細(xì)胞周期蛋白

蛋白分子量  predicted molecular weight: 43kDa

性    狀   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human P53 

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

抑制基因P53蛋白抗體（野生型P53）費(fèi)用 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 ICC=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 tumor suppressor protein containing transcriptional activation, DNA binding, and oligomerization domains. The encoded protein responds to diverse cellular stresses to regulate expression of target genes, thereby inducing cell cycle arrest, apoptosis, senescence, DNA repair, or changes in metabolism. Mutations in this gene are associated with a variety of human cancers, including hereditary cancers such as Li-Fraumeni syndrome. Alternative splicing of this gene and the use of alternate promoters result in multiple transcript variants and isoforms. Additional isoforms have also been shown to result from the use of alternate translation initiation codons (PMIDs: 12032546, 20937277). [provided by RefSeq, Feb 2013].

Function : Acts as a tumor suppressor in many tumor types; induces growth arrest or apoptosis depending on the physiological circumstances and cell type. Involved in cell cycle regulation as a trans-activator that acts to negatively regulate cell division by controlling a set of genes required for this process. One of the activated genes is an inhibitor of cyclin-dependent kinases. Apoptosis induction seems to be mediated either by stimulation of BAX and FAS antigen expression, or by repression of Bcl-2 expression. Implicated in Notch signaling cross-over. Prevents CDK7 kinase activity when associated to CAK complex in response to DNA damage, thus stopping cell cycle progression. Isoform 2 enhances the transactivation activity of isoform 1 from some but not all TP53-inducible promoters. Isoform 4 suppresses transactivation activity and impairs growth suppression mediated by isoform 1. Isoform 7 inhibits isoform 1-mediated apoptosis.

Subunit : Interacts with AXIN1. Probably part of a complex consisting of TP53, HIPK2 and AXIN1 (By similarity). Binds DNA as a homotetramer. Interacts with histone acetyltransferases EP300 and methyltransferases HRMT1L2 and CARM1, and recruits them to promoters. In vitro, the interaction of TP53 with cancer-associated/HPV (E6) viral proteins leads to ubiquitination and degradation of TP53 giving a possible model for cell growth regulation. This complex formation requires an additional factor, E6-AP, which stably associates with TP53 in the presence of E6. Interacts (via C-terminus) with TAF1; when TAF1 is part of the TFIID complex. Interacts with ING4; this interaction may be indirect. Found in a complex with CABLES1 and TP73. Interacts with HIPK1, HIPK2, and TP53INP1. Interacts with WWOX. May interact with HCV core protein. Interacts with USP7 and SYVN1. Interacts with HSP90AB1. Interacts with CHD8; leading to recruit histone H1 and prevent transactivation activity (By similarity). Interacts with ARMC10, BANP, CDKN2AIP, NUAK1, STK11/LKB1, UHRF2 and E4F1. Interacts with YWHAZ; the interaction enhances TP53 transcriptional activity. Phosphorylation of YWHAZ on 'Ser-58' inhibits this interaction. Interacts (via DNA-binding domain) with MAML1 (via N-terminus). Interacts with MKRN1. Interacts with PML (via C-terminus). Interacts with MDM2; leading to ubiquitination and proteasomal degradation of TP53. Directly interacts with FBXO42; leading to ubiquitination and degradation of TP53. Interacts (phosphorylated at Ser-15 by ATM) with the phosphatase PP2A-PPP2R5C holoenzyme; regulates stress-induced TP53-dependent inhibition of cell proliferation. Interacts with PPP2R2A. Interacts with AURKA, DAXX, BRD7 and TRIM24. Interacts (when monomethylated at Lys-382) with L3MBTL1. Isoform 1 interacts with isoform 2 and with isoform 4. Interacts with GRK5. Binds to the CAK complex (CDK7, cyclin H and MAT1) in response to DNA damage. Interacts with CDK5 in neurons. Interacts with AURKB, UHRF2 and NOC2L. Interacts (via N-terminus) with PTK2/FAK1; this promotes ubiquitination by MDM2. Interacts with PTK2B/PYK2; this promotes ubiquitination by MDM2. Interacts with PRKCG. Interacts with human cytomegalovirus/HHV-5 protein UL123.

Subcellular Location : Cytoplasm. Nucleus. Nucleus, PML body. Endoplasmic reticulum. Note=Interaction with BANP promotes nuclear localization. Recruited into PML bodies together with CHEK2.

Phosphorylated on Ser-15 upon ultraviolet irradiation; which is enhanced by interaction with BANP. Phosphorylated by NUAK1 at Ser-15 and Ser-392; was initially thought to be mediated by STK11/LKB1 but it was later shown that it is indirect and that STK11/LKB1-dependent phosphorylation is probably mediated by downstream NUAK1 (PubMed:21317932). It is unclear whether AMP directly mediates phosphorylation at Ser-15. Phosphorylated on Thr-18 by isoform 1 and isoform 2 of VRK2. Phosphorylation on Thr-18 by isoform 2 of VRK2 results in a reduction in ubiquitination by MDM2 and an increase in acetylation by EP300. Stabilized by CDK5-mediated phosphorylation in response to genotoxic and oxidative stresses at Ser-15, Ser-33 and Ser-46, leading to accumulation of p53/TP53, particularly in the nucleus, thus inducing the transactivation of p53/TP53 target genes. Phosphorylated at Ser-315 and Ser-392 by CDK2 in response to DNA-damage.

Dephosphorylated by PP2A-PPP2R5C holoenzyme at Thr-55. SV40 small T antigen inhibits the dephosphorylation by the AC form of PP2A.

May be O-glycosylated in the C-terminal basic region. Studied in EB-1 cell line.

Ubiquitinated by MDM2 and SYVN1, which leads to proteasomal degradation. Ubiquitinated by RFWD3, which works in cooperation with MDM2 and may catalyze the formation of short polyubiquitin chains on p53/TP53 that are not targeted to the proteasome. Ubiquitinated by MKRN1 at Lys-291 and Lys-292, which leads to proteasomal degradation. Deubiquitinated by USP10, leading to its stabilization. Ubiquitinated by TRIM24, which leads to proteasomal degradation. Ubiquitination by TOPORS induces degradation. Deubiquitination by USP7, leading to stabilization. Isoform 4 is monoubiquitinated in an MDM2-independent manner.

Defects in TP53 are involved in head and neck squamous cell carcinomas (HNSCC)

Defects in TP53 are a cause of lung cancer (LNCR) [MIM:211980]. LNCR is a common malignancy affecting tissues of the lung. The most common form of lung cancer is non-small cell lung cancer (NSCLC) that can be divided into 3 major histologic subtypes: squamous cell carcinoma, adenocarcinoma, and large cell lung cancer. NSCLC is often diagnosed at an advanced stage and has a poor prognosis.

Defects in TP53 are a cause of choroid plexus papilloma (CPLPA) [MIM:260500]. Choroid plexus papilloma is a slow-growing benign tumor of the choroid plexus that often invades the leptomeninges. In children it is usually in a lateral ventricle but in adults it is more often in the fourth ventricle. Hydrocephalus is common, either from obstruction or from tumor secretion of cerebrospinal fluid. If it undergoes malignant transformation it is called a choroid plexus carcinoma. Primary choroid plexus tumors are rare and usually occur in early childhood.

Defects in TP53 are a cause of adrenocortical carcinoma (ADCC) [MIM:202300]. ADCC is a rare childhood tumor of the adrenal cortex. It occurs with increased frequency in patients with the Beckwith-Wiedemann syndrome and is a component tumor in Li-Fraumeni yndrome.

Similarity : Belongs to the p53 family.

Database links : NCBI Reference Sequence: NP_001119584.1

UniProtKB/Swiss-Prot: P04637.4

wtp53廣泛的研究發(fā)現(xiàn)P53抑制基因?qū)?0%以上的人類具有抑制突變的功能。P53蛋白水平在正常細(xì)胞中表達(dá)低，在DNA突變時(shí)或各種各樣細(xì)胞遇難信號(hào)時(shí)反應(yīng)增加。該基因突變或缺失是導(dǎo)致許多發(fā)生的原因。

野生型P53(wt-p53)可誘導(dǎo)細(xì)胞凋亡，并通過細(xì)胞凋亡抑制，而P53的突變或缺失則可抑制野生型P53的功能，使得缺陷細(xì)胞得以存活下來，從而導(dǎo)致發(fā)生。

P53同時(shí)也是細(xì)胞凋亡的調(diào)控因子。此抗體可用于P53的研究。

抗體的鑒定：

1）抑制基因P53蛋白抗體（野生型P53）費(fèi)用 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?？贵w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)率來表示。交叉反應(yīng)率可用競爭抑制試驗(yàn)測定。以不同濃度抗原和近似抗原分別做競爭抑制曲線,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。

Anti-C-SKI/FITC 熒光素標(biāo)記抗C-SKI抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Anti-Laminin alpha5/FITC 熒光素標(biāo)記層粘蛋白α5抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh CCDC12 卷曲螺旋結(jié)構(gòu)域蛋白12抗體 規(guī)格 0.2ml

GITR (glucocorticoidinduced tumor necrosis factor receptor) 糖皮質(zhì)激素誘導(dǎo)壞死因子受體抗原 0.5mg

HRG1 英文名稱： 神經(jīng)調(diào)節(jié)蛋白1抗體 0.1ml

Rhesus antibody Rh SLFN14 Schlafen家族14抗體 規(guī)格 0.2ml

Anti-Laminin alpha5/FITC 熒光素標(biāo)記層粘蛋白α5抗體IgGMulti-class antibodies規(guī)格： 0.2ml

GR- Alpha ELISA Kit 大鼠糖皮質(zhì)激素受體αMulti-class antibodies規(guī)格： 48T

Anti-Phospho-TrkA (Tyr674/675)/TrkB (Tyr706/707) 磷酸化酪氨酸激酶A抗體Multi-class antibodies規(guī)格： 0.1ml

Rhesus antibody Rh GIMAP2 GTP酶IMAP家族成員2抗體 規(guī)格 0.2ml

AMA ELISA Kit 大鼠抗單核細(xì)胞抗體 96T

Phospho-NPM (Thr95) 英文名稱： 磷酸化核仁磷酸蛋白抗體 0.1ml

APPD 英文名稱： 凋亡誘導(dǎo)蛋白D抗體 0.2ml

Anti-Phospho-TrkA (Tyr674/675)/TrkB (Tyr706/707) 磷酸化酪氨酸激酶A抗體Multi-class antibodies規(guī)格： 0.1ml

Human Collagenase I ELISA Kit 人膠原酶IMulti-class antibodies規(guī)格： 48T

Anti-NFATc1 活化T細(xì)胞核因子1蛋白Multi-class antibodies規(guī)格： 0.1ml

Rhesus antibody Rh IL-2 白介素2抗體 規(guī)格 0.2ml

Col I (Mouse Collagen Type I )ELISA Kit 小鼠Ⅰ型膠原 96T

phospho-PKC beta 1/2(Thr500) 英文名稱： 磷酸化蛋白激酶C β1/β2(Thr500)抗體 0.1ml

Caveolin-1 英文名稱： 細(xì)胞質(zhì)膜微囊蛋白-1抗體 0.1ml

Anti-NFATc1 活化T細(xì)胞核因子1蛋白Multi-class antibodies規(guī)格： 0.1ml

CL-0357HHCC(人細(xì)胞)5×106cells/瓶×2

VTI1A Others Human 人 VTI1A 人細(xì)胞裂解液 (陽性對(duì)照)

人腎小球內(nèi)皮細(xì)胞總RNAHRGEC NA

綿羊皮膚細(xì)胞;OAR-S1 小鼠原成骨細(xì)胞，MC3T3-E1 Subclone 14細(xì)胞 TM3細(xì)胞，正常小鼠Leydig細(xì)胞

人皮膚成纖維細(xì)胞;HSAS4

HA Others H11N9 甲型 H11N9 (A/mallard/Alberta/294/1977) 血凝素 (Hemagglutinin / HA) 人細(xì)胞裂解液 (陽性對(duì)照)

CL-0354HFSF(人胚胎眼鞏膜成纖維細(xì)胞)5×106cells/瓶×2

NOV Others Human 人 CCN3 / NOV 桿狀病毒-昆蟲細(xì)胞裂解液 (陽性對(duì)照)

人骨骼肌衛(wèi)星細(xì)胞總RNAHSkMSC NA

CHRF細(xì)胞，人巨核細(xì)胞 人細(xì)胞系(未分化),Calu-6細(xì)胞 5637, 人細(xì)胞

家貓皮膚細(xì)胞;FCA-S1

HA Others H1N1 甲型 H1N1 (A/Brisbane/59/2007) 血凝素 (Hemagglutinin / HA) 人細(xì)胞裂解液 (陽性對(duì)照)

抑制基因P53蛋白抗體（野生型P53）費(fèi)用 BST2 Others Human 人 TETHERIN / BST2 / CD317 人細(xì)胞裂解液 (陽性對(duì)照)

雜交瘤(B類);IB3C10H12A12G2H8F3

原代上皮細(xì)胞特制無血清添加劑Many types of cells包裝：1ml

HUVE-12/CRL2480細(xì)胞，人臍內(nèi)皮細(xì)胞 長臂猿,MLA-144細(xì)胞 人真皮淋巴內(nèi)皮細(xì)胞總RNAHDLEC NA

QSG-7701(人正常肝細(xì)胞) 5×106cells/瓶×2

EL4(小鼠細(xì)胞) 5×106cells/瓶×2 CL-0255A875(人細(xì)胞)5×106cells/瓶×2 豚鼠胚胎細(xì)胞;104C1