抗体的生物素化标记实验要点：

1. 抑制基因P53蛋白抗体（野生型P53）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*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-P53 protein(wt-p53) 

中文名称   抑制基因P53蛋白抗体（野生型P53）费用 

别    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.

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, Pig, Cow, Horse, Rabbit, Sheep 

产品类型   一抗  

研究领域     细胞生物 染色质和核信号 细胞凋亡 细胞周期蛋白

蛋白分子量  predicted molecular weight: 43kDa

性    状   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human P53 

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

抑制基因P53蛋白抗体（野生型P53）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 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广泛的研究发现P53抑制基因对50%以上的人类具有抑制突变的功能。P53蛋白水平在正常细胞中表达低，在DNA突变时或各种各样细胞遇难信号时反应增加。该基因突变或缺失是导致许多发生的原因。

野生型P53(wt-p53)可诱导细胞凋亡，并通过细胞凋亡抑制，而P53的突变或缺失则可抑制野生型P53的功能，使得缺陷细胞得以存活下来，从而导致发生。

P53同时也是细胞凋亡的调控因子。此抗体可用于P53的研究。

抗体的鉴定：

1）抑制基因P53蛋白抗体（野生型P53）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。

Anti-C-SKI/FITC 荧光素标记抗C-SKI抗体IgGMulti-class antibodies规格： 0.2ml

Anti-Laminin alpha5/FITC 荧光素标记层粘蛋白α5抗体IgGMulti-class antibodies规格： 0.2ml

Rhesus antibody Rh CCDC12 卷曲螺旋结构域蛋白12抗体 规格 0.2ml

GITR (glucocorticoidinduced tumor necrosis factor receptor) 糖皮质激素诱导坏死因子受体抗原 0.5mg

HRG1 英文名称： 神经调节蛋白1抗体 0.1ml

Rhesus antibody Rh SLFN14 Schlafen家族14抗体 规格 0.2ml

Anti-Laminin alpha5/FITC 荧光素标记层粘蛋白α5抗体IgGMulti-class antibodies规格： 0.2ml

GR- Alpha ELISA Kit 大鼠糖皮质激素受体αMulti-class antibodies规格： 48T

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

Rhesus antibody Rh GIMAP2 GTP酶IMAP家族成员2抗体 规格 0.2ml

AMA ELISA Kit 大鼠抗单核细胞抗体 96T

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

APPD 英文名称： 凋亡诱导蛋白D抗体 0.2ml

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

Human Collagenase I ELISA Kit 人胶原酶IMulti-class antibodies规格： 48T

Anti-NFATc1 活化T细胞核因子1蛋白Multi-class antibodies规格： 0.1ml

Rhesus antibody Rh IL-2 白介素2抗体 规格 0.2ml

Col I (Mouse Collagen Type I )ELISA Kit 小鼠Ⅰ型胶原 96T

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

Caveolin-1 英文名称： 细胞质膜微囊蛋白-1抗体 0.1ml

Anti-NFATc1 活化T细胞核因子1蛋白Multi-class antibodies规格： 0.1ml

CL-0357HHCC(人细胞)5×106cells/瓶×2

VTI1A Others Human 人 VTI1A 人细胞裂解液 (阳性对照)

人肾小球内皮细胞总RNAHRGEC NA

绵羊皮肤细胞;OAR-S1 小鼠原成骨细胞，MC3T3-E1 Subclone 14细胞 TM3细胞，正常小鼠Leydig细胞

人皮肤成纤维细胞;HSAS4

HA Others H11N9 甲型 H11N9 (A/mallard/Alberta/294/1977) 血凝素 (Hemagglutinin / HA) 人细胞裂解液 (阳性对照)

CL-0354HFSF(人胚胎眼巩膜成纤维细胞)5×106cells/瓶×2

NOV Others Human 人 CCN3 / NOV 杆状病毒-昆虫细胞裂解液 (阳性对照)

人骨骼肌卫星细胞总RNAHSkMSC NA

CHRF细胞，人巨核细胞 人细胞系(未分化),Calu-6细胞 5637, 人细胞

家猫皮肤细胞;FCA-S1

HA Others H1N1 甲型 H1N1 (A/Brisbane/59/2007) 血凝素 (Hemagglutinin / HA) 人细胞裂解液 (阳性对照)

抑制基因P53蛋白抗体（野生型P53）费用 BST2 Others Human 人 TETHERIN / BST2 / CD317 人细胞裂解液 (阳性对照)

杂交瘤(B类);IB3C10H12A12G2H8F3

原代上皮细胞特制无血清添加剂Many types of cells包装：1ml

HUVE-12/CRL2480细胞，人脐内皮细胞 长臂猿,MLA-144细胞 人真皮淋巴内皮细胞总RNAHDLEC NA

QSG-7701(人正常肝细胞) 5×106cells/瓶×2

EL4(小鼠细胞) 5×106cells/瓶×2 CL-0255A875(人细胞)5×106cells/瓶×2 豚鼠胚胎细胞;104C1