抗體的生物素化標(biāo)記實(shí)驗(yàn)要點(diǎn)：

1. 沉默調(diào)節(jié)蛋白1抗體費(fèi)用 如在反應(yīng)混合液中有疊氮鈉或游離氨基存在,會(huì)抑制標(biāo)記反應(yīng)。因此,蛋白質(zhì)在反應(yīng)前要對(duì) 0.1mol/L碳酸氫鈉緩沖液或0.5mol/L硼酸緩沖液充分透析；

2.所用的NHSB及待生物素化蛋白質(zhì)之間的分子比按蛋白質(zhì)表面的ε-氨基的密度會(huì)有所不同,選擇不當(dāng)則影響標(biāo)記的效率,應(yīng)先用幾個(gè)不同的分子比來篩選最適條件；

3.用NHSB量過量也是不利的,抗原的結(jié)合位點(diǎn)可能因此被封閉,導(dǎo)致抗體失活；

4.由于抗體的氨基不易接近可能造成生物素化不足,此時(shí)可加入去污劑如 Triton x-100, Tween20等；

5.當(dāng)游離ε-氨基(賴氨酸殘基的氨基)存在于抗體的抗原結(jié)合位點(diǎn)時(shí),或位于酶的催化位點(diǎn)時(shí),生物素化會(huì)降低或損傷抗體蛋白的結(jié)合力或活性；

6.生物素還可能與不同的功能基團(tuán),如羰基、氨基、巰基、異咪唑基及苯酚基,也可與糖基共價(jià)結(jié)合；

7.交聯(lián)反應(yīng)后,應(yīng)充分透析,否則,殘余的生物素會(huì)對(duì)生物素化抗體與親和素的結(jié)合產(chǎn)生競爭作用；

8.在細(xì)胞的熒光標(biāo)記實(shí)驗(yàn)中,中和親和素的本底低,但由于鏈霉親和素含有少量正電荷,故對(duì)某些細(xì)胞可導(dǎo)致高本底。

產(chǎn)品訂購信息：
英文名稱  Anti-SIRT1 

中文名稱   沉默調(diào)節(jié)蛋白1抗體費(fèi)用 

別    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.

濃    度   1mg/1ml

規(guī) 格  0.1ml/100μg 0.2ml/200μg

抗體來源   Rabbit

克隆類型   polyclonal

交叉反應(yīng)   Human, Mouse, Rat, Chicken, Cow, Horse, Rabbit 

產(chǎn)品類型   一抗  

研究領(lǐng)域    細(xì)胞生物 免疫學(xué) 染色質(zhì)和核信號(hào) 微生物學(xué) 表觀遺傳學(xué)

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

性    狀   Lyophilized or Liquid

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

亞    型   IgG

純化方法   affinity purified by Protein A

儲(chǔ) 存 液   0.01M PBS, pH 7.4 with 10 mg/ml BSA and 0.1% Sodium azide

沉默調(diào)節(jié)蛋白1抗體費(fèi)用 產(chǎn)品應(yīng)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

（石蠟切片需做抗原修復(fù)） 

 not yet tested in other applications.

 optimal dilutions/concentrations should be determined by the end user.  

保存條件  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. 

產(chǎn)品介紹 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(沉默信息調(diào)節(jié)子)參與多種活動(dòng)，包括DNA的自我保護(hù)和修復(fù)，抑制脂質(zhì)過氧化積累，抑制其他細(xì)胞凋亡相關(guān)基因的表達(dá)以及和細(xì)胞壽命相關(guān)的活動(dòng)。限制攝入的熱量可以加強(qiáng)SIRT1的表達(dá)，從而延長了壽命。

SIRT蛋白成為多種生物過程的調(diào)節(jié)者也參與衰老的調(diào)控。在研究最多的SIRT蛋白中，SIRT1與各種非組蛋白或者轉(zhuǎn)錄因子相互作用，包括p53、FOXO蛋白、p300、NFkB和MyoD，sirtuins可參與凋亡、細(xì)胞存活、轉(zhuǎn)錄和代謝等過程。以sirtuins為靶標(biāo)的藥物可能在衰老、、和神經(jīng)退行性變中有用。

抗體的鑒定：

1）沉默調(diào)節(jié)蛋白1抗體費(fèi)用 抗體的效價(jià)鑒定：不管是用于診斷還是用于,制備抗體的目的都是要求較高效價(jià)。不同的抗原制備的抗體,要求的效價(jià)不一。鑒定效價(jià)的方法很多,包括有試管凝集反應(yīng),瓊脂擴(kuò)散試驗(yàn),酶聯(lián)免疫吸附試驗(yàn)等。常用的抗原所制備的抗體一般都有約成的鑒定效價(jià)的方法,以資比較。如制備抗抗體的效價(jià),一般就采用瓊脂擴(kuò)散試驗(yàn)來鑒定。

2）抗體的特異性鑒定：抗體的特異性是指與相應(yīng)抗原或近似抗原物質(zhì)的識(shí)別能力?？贵w的特異性高,它的識(shí)別能力就強(qiáng)。衡量特異性通常以交叉反應(yīng)率來表示。交叉反應(yīng)率可用競爭抑制試驗(yàn)測定。以不同濃度抗原和近似抗原分別做競爭抑制曲線,計(jì)算各自的結(jié)合率,求出各自在IC50時(shí)的濃度,并按公式計(jì)算交叉反應(yīng)率。 

如果所用抗原濃度IC50濃度為pg/管,而一些近似抗原物質(zhì)的IC50濃度幾乎是無窮大時(shí),表示這一抗血清與其他抗原物質(zhì)的交叉反應(yīng)率近似為0,即該血清的特異性較好。

3）抗體親和力：是指抗體和抗原結(jié)合的牢固程度。親和力的高低是由抗原分子的大小,抗體分子的結(jié)合位點(diǎn)與抗原決定簇之間立體構(gòu)型的合適度決定的。有助于維持抗原抗體復(fù)合物穩(wěn)定的分子間力有氫鍵,疏水鍵,側(cè)鏈相反電荷基因的庫侖力,范德華力和空間斥力。親和力常以親和常數(shù)K表示,K的單位是L/mol?？贵w親和力的測定對(duì)抗體的篩選,確定抗體的用途,驗(yàn)證抗體的均一性等均有重要意義。

Anti-Fut4/CD15/SSEA-1 /FITC 熒光素標(biāo)記階段特異性胚胎表面抗原-1抗體IgGMulti-class antibodies規(guī)格： 0.2ml

GAP-43 ( Growth associated protein-43 ) 生長相關(guān)蛋白-43(抗原)Multi-class antibodies規(guī)格： 0.5mg

血小板源性生長因子-BB抗體 Anti-PDGF-BB 0.2ml

SLCO1C1 英文名稱： 溶質(zhì)載體家族蛋白21成員1C1抗體 0.1ml

ELAVL1/HuR 英文名稱： ELAVL1抗體 0.2ml

Rhesus antibody Rh Phospho-NDRG1 (Ser330) 磷酸化分化相關(guān)基因NDRG1抗體 規(guī)格 0.2ml

GAP-43 ( Growth associated protein-43 ) 生長相關(guān)蛋白-43(抗原)Multi-class antibodies規(guī)格： 0.5mg

Anti-TNF- Beta/FITC 熒光素標(biāo)記壞死因子-β抗體IgGMulti-class antibodies規(guī)格： 0.2ml

PDGF-B 血小板源性生長因子-B(抗原)Multi-class antibodies規(guī)格： 0.5mg

金屬蛋白酶組織抑制因子-3抗體 Anti-TIMP-3 0.1ml

5HT4 Receptor 英文名稱： 5-羥色胺受體4抗體 0.1ml

ETHE1 英文名稱： 乙基腦病蛋白抗體 0.2ml

Rhesus antibody Rh phospho-KCND2/Kv4.2(Thr607) 磷酸化電壓門控性通道蛋白Kv4.2抗體 規(guī)格 0.1ml

PDGF-B 血小板源性生長因子-B(抗原)Multi-class antibodies規(guī)格： 0.5mg

PDGF-BB peptide 血小板源性生長因子-BB抗原Multi-class antibodies規(guī)格： 0.5mg

Anti-ATP5J ATP5J抗體Multi-class antibodies規(guī)格： 0.1ml

Rhesus antibody Rh phospho-ANAPC1 (Ser355) 磷酸化細(xì)胞周期末期促進(jìn)復(fù)合蛋白APC1抗體 規(guī)格 0.1ml

p27 濃縮液 0.1ml 進(jìn)口分裝

UCP-3 英文名稱： 線粒體脫偶連蛋白3抗體 0.1ml

Defensin beta 2 英文名稱： 防御素β2/Defensin β2抗體 0.1ml

Anti-ATP5J ATP5J抗體Multi-class antibodies規(guī)格： 0.1ml

IL23 Protein Human 重組人 IL-23 (IL23A & IL12B Heterodimer) 蛋白

小鼠真皮成纖維細(xì)胞完全培養(yǎng)基 100mL

LRP10 Others Human 人 LRP10 人細(xì)胞裂解液 (陽性對(duì)照)

HN13口腔細(xì)胞 HN13 oral squamous cell carcinoma cells DMEM+10%FBS

CM-R100大鼠腦動(dòng)脈內(nèi)皮細(xì)胞完全培養(yǎng)基100mL

UM-UC-3(人膀胱移行細(xì)胞癌) 5×106cells/瓶×2 HUM, 正常人主動(dòng)脈平滑肌細(xì)胞

CM-M010小鼠肺大動(dòng)脈平滑肌細(xì)胞完全培養(yǎng)基100mL

KIAA1279 Others Human 人 KIAA1279 桿狀病毒-昆蟲細(xì)胞裂解液 (陽性對(duì)照)

腎系膜細(xì)胞培養(yǎng)基MCM

SV40轉(zhuǎn)化的非洲綠猴腎細(xì)胞;COS-7 細(xì)胞，Jecket細(xì)胞 A172(膠質(zhì)母細(xì)胞瘤細(xì)胞)

EB病毒轉(zhuǎn)化的人B淋巴細(xì)胞;KMY0907

ACVR2A Others Mouse 小鼠 ACVR2A / AcIIa 人細(xì)胞裂解液 (陽性對(duì)照)

沉默調(diào)節(jié)蛋白1抗體費(fèi)用 IL23 Protein Human 重組人 IL-23 (IL23A & IL12B Heterodimer) 蛋白

小鼠真皮成纖維細(xì)胞完全培養(yǎng)基 100mL

LRP10 Others Human 人 LRP10 人細(xì)胞裂解液 (陽性對(duì)照)

HN13口腔細(xì)胞 HN13 oral squamous cell carcinoma cells DMEM+10%FBS

CM-R100大鼠腦動(dòng)脈內(nèi)皮細(xì)胞完全培養(yǎng)基100mL

UM-UC-3(人膀胱移行細(xì)胞癌) 5×106cells/瓶×2 HUM, 正常人主動(dòng)脈平滑肌細(xì)胞