抗体的生物素化标记实验要点：

1. 丝氨酸/苏氨酸蛋白激酶PIM3抗体品牌 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-PIM3 

中文名称   丝氨酸/苏氨酸蛋白激酶PIM3抗体品牌 

别    名  Kid 1; Kid-1; Kid1; Kinase induced by depolarization; ONCOGENE PIM 3; ONCOGENE PIM3; PIM 3; Pim 3 oncogene; PIM3; Pim3 oncogene; PIM3_HUMAN; Protein kinase Kid 1; Protein kinase Kid1; Serine/threonine kinase Pim 3; Serine/threonine kinase Pim3; Serine/threonine protein kinase Pim 3; Serine/threonine protein kinase Pim3; Serine/threonine-protein kinase Pim-3.

浓    度   1mg/1ml

规 格  0.2ml/200μg

抗体来源   Rabbit

克隆类型   polyclonal

交叉反应   Human, Mouse, Rat, Chicken, Dog, Pig, Cow, Rabbit, Sheep, Guinea Pig 

产品类型   一抗  

研究领域     细胞生物 信号转导 激酶和磷酸酶

蛋白分子量  predicted molecular weight: 36kDa

性    状   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human PIM3 

亚    型   IgG

纯化方法   affinity purified by Protein A

储 存 液   Preservative: 15mM Sodium Azide, Constituents: 1% BSA, 0.01M PBS, pH 7.4

丝氨酸/苏氨酸蛋白激酶PIM3抗体品牌 产品应用    WB=1:100-500 ELISA=1:500-1000 IHC-P=1:100-500 IHC-F=1:100-500 IF=1:50-200

（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 The Pim family serine/threonine protein kinases were first identified in studies examining genes targeted for proviral insertion in murine leukemia virus-induced T lymphomas. Increased levels of Pim kinases predispose cells to lymphomagenesis and enhance the activity of mitogenic proteins such as p100, c-Myb, and Cdc25A. In addition, Pim kinases are also involved in modulation of synaptic strength in neurons and anti-apoptotic signaling in hematopoietic progenitor cells. Pim-3, a member of the proto-oncogene Pim family that expresses serine/threonine kinase activity, shares significant homology with Pim-1 serine/threonine protein kinases. Pim-3 may function as a mediator of synaptic plasticity in the brain and is presumably involved in the anti-apoptosis process and cell cycle progression as well as the proliferation of human hepatoma cell lines. The Pim-3 protein is widely expressed, however no expression is observed in the colon, thymus, or small intestine.

Function : Proto-oncogene with serine/threonine kinase activity that can prevent apoptosis, promote cell survival and protein translation. May contribute to tumorigenesis through: the delivery of survival signaling through phosphorylation of BAD which induces release of the anti-apoptotic protein Bcl-X(L), the regulation of cell cycle progression, protein synthesis and by regulation of MYC transcriptional activity. Additionally to this role on tumorigenesis, can also negatively regulate insulin secretion by inhibiting the activation of MAPK1/3 (ERK1/2), through SOCS6. Involved also in the control of energy metabolism and regulation of AMPK activity in modulating MYC and PPARGC1A protein levels and cell growth.

Subunit : Interacts with BAD. Interacts with PPP2CA; this interaction promotes dephosphorylation of PIM3, ubiquitination and proteasomal degradation. Interacts with SOCS6 (By similarity).

Subcellular Location : Cytoplasm.

Post-translational modifications : Ubiquitinated, leading to proteasomal degradation.

Phosphorylated. Interaction with PPP2CA promotes dephosphorylation.

Similarity : Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. PIM subfamily.

Contains 1 protein kinase domain.

Database links :

UniProtKB/Swiss-Prot: Q86V86.3

Entrez Gene: 415116 Human

Entrez Gene: 223775 Mouse

Omim: 610580 Human

SwissProt: Q86V86 Human

SwissProt: P58750 Mouse

Unigene: 530381 Human

Unigene: Hs.530381 Human

Unigene: 400129 Mouse

抗体的鉴定：

1）丝氨酸/苏氨酸蛋白激酶PIM3抗体品牌 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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