抗体的生物素化标记实验要点：

1. S100钙结合蛋白A6抗体规格 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、异咪唑基及苯酚基,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-S100A6 

中文名称   S100钙结合蛋白A6抗体规格 

别    名  S100 alpha 6 PRA; Calcyclin; PRAGrowth factor inducible protein 2A9; Prolactin receptor associated protein; Prolactin receptor-associated protein; Protein S100 A6; Protein S100-A6; S100 A6; S100 calcium binding protein A6 (calcyclin); S100 calcium binding protein A6; S100 calcium-binding protein A6; S100A6; S100A6_HUMAN; S100-A6; 2A9; 5B10; CABP; CACY; Calcyclin; Growth factor inducible protein 2A9; Growth factor-inducible protein 2A9; MLN 4; MLN4.

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, Dog, Pig, Horse, Rabbit 

产品类型   一抗  

研究领域    免疫学 信号转导 生长因子和激素 细胞因子

蛋白分子量  predicted molecular weight: 10kDa

性    状   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human S100A6 

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

S100钙结合蛋白A6抗体规格 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 Calcyclin, also known as Protein S100-A6, growth factor-inducible protein 2A9, S-100 calcium-binding protein A6 (S-100A6) or MLN 4, is a homodimeric member of the S-100 calcium-binding protein family whose expression is upregulated in proliferating and differentiating cells. Calcyclin is inducible by growth factors and overexpressed in acute myeloid leukemias. It is expressed in a cell-specific manner in subpopulations of neurons and astrocytes and in epithelial cells and fibroblasts. Calcyclin is a specific target of S-100B protein in vivo. The binding of Calcyclin to S-100B is stabilized by S-100B-bound calcium and zinc. Calcyclin associates with both Annexin XI and CacyBP (calcyclin-binding protein). It functions to activate several processes along the calcium signal transduction pathway including the regulation of cell growth, proliferation, secretion and exocytosis.

Function : May function as calcium sensor and contribute to cellular calcium signaling (Potential). May function by interacting with other proteins and indirectly play a role in the reorganization of the actin cytoskeleton and in cell motility. Binds 2 calcium ions. Calcium binding is cooperative.

Subunit : Homodimer; head to tail assembly of 2 subunits. Interacts with CACYBP. Interacts with ANXA2 and ANXA11 (via N-terminus). Interacts with SUGT1. Interacts with TP53; has higher affinity for TP53 that is phosphorylated on its N-terminal domain, and lower affinity for TP53 that is phosphorylated on its C-terminal domain. Interacts with tropomyosin. Interacts with FKBP4. Interacts with CACYBP in a calcium-dependent manne.

Subcellular Location : Nucleus envelope. Cytoplasm. Cell membrane; Peripheral membrane protein; Cytoplasmic side.

Post-translational modifications : The N-terminus is blocked.

Similarity : Belongs to the S-100 family.Contains 2 EF-hand

Database links : UniProtKB/Swiss-Prot: P06703.1

抗体的鉴定：

1）S100钙结合蛋白A6抗体规格 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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TrK A 浓缩液 0.1ml 进口分装

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6-10B, 人细胞系

3T3 小鼠成纤维细胞

FCGR2B Others Cynomolgus 食蟹猴 CD32b / FCGR2B 人细胞裂解液 (阳性对照)

中国仓鼠卵巢细胞(亚系克隆);CHO-HCV-NS4A

甲状腺滤泡上皮细胞Many types of cells包装：5 × 105次方(1ml)

人神经母细胞瘤细胞;BE(2)-M17 大鼠肝动脉平滑肌细胞完全培养基 100mL

5637, 人细胞

SERPINB10 Others Mouse 小鼠 SerpinB10 杆状病毒-昆虫细胞裂解液 (阳性对照)

腺病毒转化的人胚肾细胞;AAV-293

HCCLM3细胞，细胞 人细胞系,HNE2-LMP1/2细胞 CL-0354HFSF(人胚胎眼巩膜成纤维细胞)5×106cells/瓶×2

中国仓鼠卵巢细胞(亚系克隆);CHO Hu 164 SCF 17

LCN2 Others Mouse 小鼠 LCN2 / NGAL 人细胞裂解液 (阳性对照)

S100钙结合蛋白A6抗体规格 6-10B, 人细胞系

3T3 小鼠成纤维细胞

FCGR2B Others Cynomolgus 食蟹猴 CD32b / FCGR2B 人细胞裂解液 (阳性对照)

中国仓鼠卵巢细胞(亚系克隆);CHO-HCV-NS4A

甲状腺滤泡上皮细胞Many types of cells包装：5 × 105次方(1ml)

人神经母细胞瘤细胞;BE(2)-M17 大鼠肝动脉平滑肌细胞完全培养基 100mL