Gene : | TP63 |
Gene ID : | 8626 |
Uniprot ID : | Q9H3D4 |
Alternative Name : | Chronic ulcerative stomatitis protein, Keratinocyte transcription factor KET, Transformation-related protein 63, Tumor protein p73-like, p40, p51, KET, P63, P73H, P73L, TP73L |
Amount : | 0.5 ml |
Isotype : | Mouse IgG1 |
The p63 gene is a homologue of the p53 tumor suppressor gene. Like p53, p63 contains a transactivation (TA) domain induce the transcription of target genes, a DNA binding domain, and an oligomerization domain (OD), used to form tetramers. In contrast to p53, the p63 gene encodes for at least six major isotypes. Three isotypes (TAp63α, TAp63β, and TAp63γ) contain the transactivating (TA) domain and are able to transactivate p53 report genes and induce apoptosis. In contrast, the other three isotypes (ΔNp63α, ΔNp63β, ΔNp63γ) are transcribed from an internal promoter localized within intron3, lack the TA domain, and act as dominant-negatives to suppress transactivation by both p53 and TAp63 isotypes. p63 is highly expressed in the basal cells of the epithelium significant for proper limb outgrowth and morphogenesis. In differentiating tissues, p63 is crucial for maintaining the stem cell identity of the basal cells, and is indispensable for correct development of the skin as well as the limb. p63-deficient mice lack all squamous epithelia and their derivatives, including hair, whiskers, teeth, as well as mammary, lacrimal, and salivary glands.Tissue specificity: Widely expressed, notably in heart, kidney, placenta, prostate, skeletal muscle, testis and thymus, although the precise isoform varies according to tissue type. Progenitor cell layers of skin, breast, eye and prostate express high levels of DeltaN-type isoforms. Isoform 10 is predominantly expressed in skin squamous cell carcinomas, but not in normal skin tissues.
Content : | TRIS with 0.03% sodium azide, pH7.2 |
Storage condition : | Store at 4°C |
Immunohistochemical Analysis :-1:250
For Research Use Only. Not for use in diagnostic/therapeutics procedures.
Subcellular location: | Nucleus |
Post transnational modification: | Ubiquitinated. Polyubiquitination involves WWP1 and leads to proteasomal degradation of this protein. |
Tissue Specificity: | Widely expressed, notably in heart, kidney, placenta, prostate, skeletal muscle, testis and thymus, although the precise isoform varies according to tissue type. Progenitor cell layers of skin, breast, eye and prostate express high levels of DeltaN-type isoforms. Isoform 10 is predominantly expressed in skin squamous cell carcinomas, but not in normal skin tissues. |
BioGrid: | 114181.129 interactions. |
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荧光燃料使用的问题有两个:
一个就是迁移性,使用时要严格控制用量,以避免产生迁移现象;
二是荧光染料使用有一个极限值,如果超量使用,导致荧光线被吸收,使吸收和发光成叠所致。
想请问一下,DAPI这个染料到底有没有膜通透性,我通过百度搜索查询关于DAPI染料的,基本上是说它能透过细胞膜对活细胞和死细胞均能染上蓝色;但是也有人说DAPI只可以透过死细胞膜,不能对活细胞进行染色,用以区分活死细胞,到底哪个是对的啊,蒙了!!!!!!
抗原抗体反应后,利用特殊仪器测定荧光强度而推算被测物浓度的检测方法
⑴荧光物质
1)荧光色素
许多物质都可产生荧光现象,但并非都可用作荧光色素。只有那些能产生明显的荧光并能作为染料使用的有机化合物才能称为免疫荧光色素或荧光染料。常用的荧光色素有:
⑴异硫氰酸荧光素(fluoresceinisothiocyanate,FITC)为黄色或橙黄色结晶粉末,易溶于水或酒精等溶剂。分子量为389.4,最大吸收光波长为490--495nm,最大发射光波长520--530nm,呈现明亮的黄绿色荧光,结构式如下:
有两种同分异结构,其中异构体Ⅰ型在效率、稳定性、与蛋白质结合能力等方面都更好,在冷暗干燥处可保存多年,是应用最广泛的荧光素。其主要优点是:①人眼对黄绿色较为敏感,②通常切片标本中的绿色荧光少于红色。
⑵四乙基罗丹明(rhodamine,RIB200)为橘红色粉末,不溶于水,易溶于酒精和丙酮。性质稳定,可长期保存。结构式如下:
最大吸收光波长为570nm,最大发射光波长为595~600nm,呈橘红色荧光。
⑶四甲基异硫氰酸罗丹明(tetramethylrhodamineisothiocyanate,TRITC)结构式如下:
最大吸引光波长为550nm,最大发射光波长为620nm,呈橙红色荧光。与FITC的翠绿色荧光对比鲜明,可配合用于双重标记或对比染色。其异硫氰基可与蛋白质结合,但荧光效率较低。
⑵其他荧光物质
1)酶作用后产生荧光的物质某些化合物本身无荧光效应,一旦经酶作用便形成具有强荧光的物质。例如4-甲基伞酮-β-D半乳糖苷受β-半乳糖苷酶的作用分解成4-甲基伞酮,后者可发出荧光,激发光波长为360nm,发射光波长为450nm。其他如碱性酸酶的底物4-甲基伞酮磷酸盐和辣根过氧化物酶的底物对羟基苯乙酸等。
2)镧系螯合物某些3价稀土镧系元素如铕(Eu3 )、铽(Tb3 )、铈(Ce3 )等的螯合物经激发后也可发射特征性的荧光,其中以Eu3 应用最广。Eu3
螯合物的激发光波长范围宽,发射光波长范围窄,荧光衰变时间长,最适合用于分辨荧光免疫测定。展开
这些染料都非常成熟,光毒和淬灭都很低,当然要考虑到你采集图像时的显微镜参数。
比如calcein,常用的示踪,绿光(虽然这些颜色只是根据光谱加上去的伪彩),但要考虑你的实验过程中结合细胞结构,是否会伴随calcein的泄漏,就是荧光降低。
dri,还可以看膜啊
cfda也不错
bcef虽是ph指示,但你试验时不仅可观察细胞,还可看细胞ph变化,也行。
当然你或许还要结合其他方法,如细胞免疫化学等手段去双染或多染,都需要综合考虑染料之间特性。单独染一个染料,有点浪费,不如多染,数据和图像也好看些。现在流行细胞成像。
我想用共聚焦观察间期染色体,用涂染探针,有一个问题很困扰我,我想涂染完染色后用DAPI复染细胞核,但是目前哈尔滨的共聚焦都没有紫外激发光,不能激发DAPI,我怎么才能实现,用红色涂一条染色体,用绿色涂另一条,用DAPI蓝色染核,同时成像呢,所说的双光子显微镜可以么?我实在很迷惑,求求版主别再删我的帖子,尽管我现在只是一个索取者,还不能给大家提供有用的信息,但是相信有一天会为大家做贡献的,谢谢了
1.小鼠Lewis肺癌细胞DNA含量测定方法
(1).从C57BL/6小鼠上切除肿块,在培养皿内用PBS冲洗.
(2).去除结缔组织及脂肪,剪碎肿块.
(3).小碎片移入1.20×38mm注射针,加压使其通过,于4℃条件下重悬细胞于HBSS中.
(4).将200~300μL细胞悬液(5×105细胞/mL)中加入3mL PI(50μg/mL),染色3LL细胞,于4℃存放20~30分钟.
(5).测定580~750nm之间的发射荧光,以去除末结合PI产生的激发光与发射光谱线之间的重叠部分.
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