抗体的生物素化标记实验要点：

1. 监理蛋白抗体品牌 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-Supervillin 

中文名称   监理蛋白抗体品牌 

别    名  Archvillin; p205/p250; SVIL_HUMAN.

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, Chicken, Dog, Pig, Cow, Horse, Sheep 

产品类型   一抗  

研究领域    细胞生物 信号转导 表观遗传学

蛋白分子量  predicted molecular weight: 248kDa

性    状   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human Supervillin 

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

监理蛋白抗体品牌 产品应用    ELISA=1:500-1000 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 Supervillin is an F-actin-binding protein originally with two reported isoforms of 205/250 kDa. The longer isoform, named archvillin, is muscle-specific. Supervillin is a tightly bound peripheral membrane protein that is concentrated at sites of epithelial cell-cell adhesion. It contributes to cell-cell adhesion, motility regulation, and information transfer between cell compartments. The COOH-terminus of supervillin is homologous to villin/gelsolin but is not responsible for the tight binding to the actin cytoskeleton in vivo. The NH2-terminus contains functional nuclear localization sequences and F-actin and myosin II binding domains. It has been suggested that supervillin may mediate actin and myosin II filament organization at cholesterol-rich membrane domains. Supervillin has been identified as a transcriptional activator of the androgen receptor. Supervillin is found in many cells of several species, but is most abundant in muscle, bone marrow, thyroid gland, and salivary gland.

Function : Forms a high-affinity link between the actin cytoskeleton and the membrane. Isoform 1 (archvillin) is among the first costameric proteins to assemble during myogenesis and it contributes to myogenic membrane structure and differentiation. Appears to be involved in myosin II assembly. May modulate myosin II regulation through MLCK during cell spreading, an initial step in cell migration. May play a role in invadopodial function. Isoform 2 may be involved in modulation of focal adhesions. Supervillin-mediated down-regulation of focal adehesions involves binding to TRIP6 (By similarity).

Subunit : Associates with F-actin. Interacts with NEB. Interacts with MYH9. Interacts with MYLK. Isoform 2 interacts with TRIP6. Isoform 2 interacts with DYNLT1 (By similarity).

Subcellular Location : Cell membrane; peripheral membrane protein; cytoplasmic side. Cytoplasm; cytoskeleton.

Tissue Specificity : Expressed in many tissues. Most abundant in muscle, bone marrow, thyroid gland and salivary gland. Isoform 1 (archvillin) is muscle specific.

Similarity : Belongs to the villin/gelsolin family.

Contains 5 gelsolin-like repeats.

Contains 1 HP (headpiece) domain.

Database links : UniProtKB/Swiss-Prot: O95425.2

抗体的鉴定：

1）监理蛋白抗体品牌 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。

ROCK2 (Rho-associated coiled-coil containing protein kinase 2) Rho相关蛋白激酶2抗原Multi-class antibodies规格： 0.5mg

Anti-Phospho-Bad(Ser136) 磷酸化相关死亡促进因子抗体Multi-class antibodies规格： 0.1ml

Rhesus antibody Rh phgophs-ATG4C(Ser451) 磷酸化自噬相关蛋白4C抗体 规格 0.1ml

Kappa Light Chain 浓缩液 0.1ml 进口分装

Urokinase 英文名称： 尿激酶型纤溶酶原激活因子抗体 0.1ml

C4b-A 英文名称： 补体C4b-A蛋白抗体 0.1ml

Anti-Phospho-Bad(Ser136) 磷酸化相关死亡促进因子抗体Multi-class antibodies规格： 0.1ml

Anti-PPAR-delta /FITC 荧光素标记D型-过氧化酶活化增生受体抗体IgGMulti-class antibodies规格： 0.2ml

Rabbit Anti-Biotin Whole serum 兔抗生物素抗血清Multi-class antibodies规格： 1ml

干扰素-γ抗体 Anti-IFN-γAnti-Human 0.1ml

Donkey Anti-Goat IgG 驴抗羊IgG 1mg

FAM48A 英文名称： P38相互作用蛋白抗体 0.1ml

Rhesus antibody Rh phospho-RAD51(Tyr315) 磷酸化Rad51抗体 规格 0.1ml

Rabbit Anti-Biotin Whole serum 兔抗生物素抗血清Multi-class antibodies规格： 1ml

Phospho-Smad3 (Ser208) 英文名称： 磷酸化细胞信号转导分子SMAD3抗体 0.1ml

Phospho-Ezrin (Tyr353) 英文名称： 磷酸化埃兹蛋白抗体 0.1ml

三叶肽因子3抗体 Anti-TFF3 0.1ml

Anti-TLR2/CD282 /FITC 荧光素标记Toll样受体2抗体IgGMulti-class antibodies规格： 0.2ml

Rhesus antibody Rh phospho-LEF1(Ser42) 磷酸化淋巴增强因子-1抗体 规格 0.1ml

phospha-Bcl-xL (pSer62) peptide (human, mo, rat, bovine, dog, pig, sheep, horse) 磷酸化(Ser62)Bcl-xL蛋白抗原Multi-class antibodies规格： 0.5mg

HL-60, 人早幼粒细胞

B16 小鼠细胞

FCGR3A Others Human 人 CD16a / FCGR3A 人细胞裂解液 (阳性对照)

大鼠肾小球系膜细胞;HBZY-1

肝动脉平滑肌细胞Many types of cells包装：5 × 105次方(1ml)

人细胞;SF17 大鼠肝窦内皮细胞完全培养基 100mL

293T, SV40转化的人胚肾上皮细胞系

KLK11 Others Mouse 小鼠 KLK11 / Kallikrein-11 人细胞裂解液 (阳性对照)

&CHAR(10)&

NIH-3T3 小鼠成纤维细胞 NIH-3T3 mouse fibroblast cells DMEM+10% FBS

IFNL1 Protein Human 重组人 IL-29 / Ierleukin-29 蛋白 (His 标签)

NCI-H1703(人肺鳞癌细胞) 5×106cells/瓶×2 PA317(成纤维细胞)

监理蛋白抗体品牌 HL-60, 人早幼粒细胞

B16 小鼠细胞

FCGR3A Others Human 人 CD16a / FCGR3A 人细胞裂解液 (阳性对照)

大鼠肾小球系膜细胞;HBZY-1

肝动脉平滑肌细胞Many types of cells包装：5 × 105次方(1ml)

人细胞;SF17 大鼠肝窦内皮细胞完全培养基 100mL