抗体的生物素化标记实验要点：

1. 沉默调节蛋白1抗体费用 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-SIRT1 

中文名称   沉默调节蛋白1抗体费用 

别    名  75SirT1; BA57G10.4; hSIR2; hSIRT1; NAD dependent deacetylase SIRT1; NAD dependent deacetylase sirtuin 1; NAD dependent deacetylase sirtuin 2; Silent mating type information regulation 2; OTTHUMP00000198111; OTTHUMP00000198112; SIR2 like 1; SIR2 like protein 1; SIR2-like protein 1; Regulatory protein SIR2 homolog 1; SIR1_HUMAN; SIR2ALPHA; SIR2alpha protein; SIR2L1; SIR2L2; SIRT 1; SIRT-1; Sirt1; SIRT1 Sir2 like proteins (siruitins) type 1; SIRT1: sirtuin (silent mating type information regulation 2 homolog) 1 (S. cerevisiae); sirtuin (silent mating type information regulation 2 homolog) 1 (S. cerevisiae); SirtT1 75 kDa fragment; sirtuin (silent mating type information regulation 2 homolog) 1 (S. cerevisiae);Sirtuin 1; sirtuin; Sirtuin type 1; Sirtuin type 2.

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, Cow, Horse, Rabbit 

产品类型   一抗  

研究领域    细胞生物 免疫学 染色质和核信号 微生物学 表观遗传学

蛋白分子量  predicted molecular weight: 58/81kDa

性    状   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human SIRT1 C-terminus 

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

沉默调节蛋白1抗体费用 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 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 member of the sirtuin family of proteins, homologs to the yeast Sir2 protein. Members of the sirtuin family are characterized by a sirtuin core domain and grouped into four classes. The functions of human sirtuins have not yet been determined; however, yeast sirtuin proteins are known to regulate epigenetic gene silencing and suppress recombination of rDNA. Studies suggest that the human sirtuins may function as intracellular regulatory proteins with mono-ADP-ribosyltransferase activity. The protein encoded by this gene is included in class I of the sirtuin family. Alternative splicing results in multiple transcript variants.

Function : SirtT1 75 kDa fragment: catalytically inactive 75SirT1 may be involved in regulation of apoptosis. May be involved in protecting chondrocytes from apoptotic death by associating with cytochrome C and interfering with apoptosome assembly.

Subunit : Found in a complex with PCAF and MYOD1 (By similarity). Component of the eNoSC complex, composed of SIRT1, SUV39H1 and RRP8. Interacts with HES1, HEY2 and PML. Interacts with RPS19BP1/AROS. Interacts with KIAA1967/DBC1 (via N-terminus); the interaction disrupts the interaction between SIRT1 and p53/TP53. Interacts with SETD7; the interaction induces the dissociation of SIRT1 from p53/TP53 and increases p53/TP53 activity. Interacts with MYCN, NR1I2, CREBZF, TSC2, TLE1, FOS, JUN, NR0B2, PPARG, NCOR, IRS1, IRS2 and NMNAT1. Interacts with HNF1A; the interaction occurs under nutrient restriction. Interacts with SUZ12; the interaction mediates the association with the PRC4 histone methylation complex which is specific as an association with PCR2 and PCR3 complex variants is not found. Interacts with -1 tat.

Subcellular Location : Nucleus, PML body. Cytoplasm. Note=Recruited to the nuclear bodies via its interaction with PML. Colocalized with APEX1 in the nucleus. May be found in nucleolus, nuclear euchromatin, heterochromatin and inner membrane. Shuttles between nucleus and cytoplasm.

SirtT1 75 kDa fragment: Cytoplasm. Mitochondrion.

Tissue Specificity : Widely expressed.

Post-translational modifications : Methylated on multiple lysine residues; methylation is enhanced after DNA damage and is dispensable for deacetylase activity toward p53/TP53.

Phosphorylated. Phosphorylated by STK4/MST1, resulting in inhibition of SIRT1-mediated p53/TP53 deacetylation. Phosphorylation by MAPK8/JNK1 at Ser-27, Ser-47, and Thr-530 leads to increased nuclear localization and enzymatic activity. Phosphorylation at Thr-530 by DYRK1A and DYRK3 acivates deacetylase activity and promotes cell survival. Phosphorylation by mammalian target of rapamycin complex 1 (mTORC1) at Ser-47 inhibits deacetylation activity. Phosphorylated by CaMK2, leading to increased p53/TP53 and NF-kappa-B p65/RELA deacetylation activity (By similarity). Phosphorylation at Ser-27 implicating MAPK9 is linked to protein stability. There is some ambiguity for some phosphosites: Ser-159/Ser-162 and Thr-544/Ser-545.

Proteolytically cleaved by cathepsin B upon TNF-alpha treatment to yield catalytic inactive but stable SirtT1 75 kDa fragment (75SirT1).

S-nitrosylated by GAPDH, leading to inhibit the NAD-dependent protein deacetylase activity (By similarity).

Similarity : Belongs to the sirtuin family.

Contains 1 deacetylase sirtuin-type domain.

Database links : UniProtKB/Swiss-Prot: Q96EB6.2

在Sirtuin蛋白家族中，sirtuin 1(沉默信息调节子)参与多种活动，包括DNA的自我保护和修复，抑制脂质过氧化积累，抑制其他细胞凋亡相关基因的表达以及和细胞寿命相关的活动。限制摄入的热量可以加强SIRT1的表达，从而延长了寿命。

SIRT蛋白成为多种生物过程的调节者也参与衰老的调控。在研究最多的SIRT蛋白中，SIRT1与各种非组蛋白或者转录因子相互作用，包括p53、FOXO蛋白、p300、NFkB和MyoD，sirtuins可参与凋亡、细胞存活、转录和代谢等过程。以sirtuins为靶标的药物可能在衰老、、和神经退行性变中有用。

抗体的鉴定：

1）沉默调节蛋白1抗体费用 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-Fut4/CD15/SSEA-1 /FITC 荧光素标记阶段特异性胚胎表面抗原-1抗体IgGMulti-class antibodies规格： 0.2ml

GAP-43 ( Growth associated protein-43 ) 生长相关蛋白-43(抗原)Multi-class antibodies规格： 0.5mg

血小板源性生长因子-BB抗体 Anti-PDGF-BB 0.2ml

SLCO1C1 英文名称： 溶质载体家族蛋白21成员1C1抗体 0.1ml

ELAVL1/HuR 英文名称： ELAVL1抗体 0.2ml

Rhesus antibody Rh Phospho-NDRG1 (Ser330) 磷酸化分化相关基因NDRG1抗体 规格 0.2ml

GAP-43 ( Growth associated protein-43 ) 生长相关蛋白-43(抗原)Multi-class antibodies规格： 0.5mg

Anti-TNF- Beta/FITC 荧光素标记坏死因子-β抗体IgGMulti-class antibodies规格： 0.2ml

PDGF-B 血小板源性生长因子-B(抗原)Multi-class antibodies规格： 0.5mg

金属蛋白酶组织抑制因子-3抗体 Anti-TIMP-3 0.1ml

5HT4 Receptor 英文名称： 5-羟色胺受体4抗体 0.1ml

ETHE1 英文名称： 乙基脑病蛋白抗体 0.2ml

Rhesus antibody Rh phospho-KCND2/Kv4.2(Thr607) 磷酸化电压门控性通道蛋白Kv4.2抗体 规格 0.1ml

PDGF-B 血小板源性生长因子-B(抗原)Multi-class antibodies规格： 0.5mg

PDGF-BB peptide 血小板源性生长因子-BB抗原Multi-class antibodies规格： 0.5mg

Anti-ATP5J ATP5J抗体Multi-class antibodies规格： 0.1ml

Rhesus antibody Rh phospho-ANAPC1 (Ser355) 磷酸化细胞周期末期促进复合蛋白APC1抗体 规格 0.1ml

p27 浓缩液 0.1ml 进口分装

UCP-3 英文名称： 线粒体脱偶连蛋白3抗体 0.1ml

Defensin beta 2 英文名称： 防御素β2/Defensin β2抗体 0.1ml

Anti-ATP5J ATP5J抗体Multi-class antibodies规格： 0.1ml

IL23 Protein Human 重组人 IL-23 (IL23A & IL12B Heterodimer) 蛋白

小鼠真皮成纤维细胞完全培养基 100mL

LRP10 Others Human 人 LRP10 人细胞裂解液 (阳性对照)

HN13口腔细胞 HN13 oral squamous cell carcinoma cells DMEM+10%FBS

CM-R100大鼠脑动脉内皮细胞完全培养基100mL

UM-UC-3(人膀胱移行细胞癌) 5×106cells/瓶×2 HUM, 正常人主动脉平滑肌细胞

CM-M010小鼠肺大动脉平滑肌细胞完全培养基100mL

KIAA1279 Others Human 人 KIAA1279 杆状病毒-昆虫细胞裂解液 (阳性对照)

肾系膜细胞培养基MCM

SV40转化的非洲绿猴肾细胞;COS-7 细胞，Jecket细胞 A172(胶质母细胞瘤细胞)

EB病毒转化的人B淋巴细胞;KMY0907

ACVR2A Others Mouse 小鼠 ACVR2A / AcIIa 人细胞裂解液 (阳性对照)

沉默调节蛋白1抗体费用 IL23 Protein Human 重组人 IL-23 (IL23A & IL12B Heterodimer) 蛋白

小鼠真皮成纤维细胞完全培养基 100mL

LRP10 Others Human 人 LRP10 人细胞裂解液 (阳性对照)

HN13口腔细胞 HN13 oral squamous cell carcinoma cells DMEM+10%FBS

CM-R100大鼠脑动脉内皮细胞完全培养基100mL

UM-UC-3(人膀胱移行细胞癌) 5×106cells/瓶×2 HUM, 正常人主动脉平滑肌细胞