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Mouse anti DDDDK-Tag mAb (AE005)

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Western blot analysis of lysates from wild type (WT) and 293F cells transfected with Mouse anti DDDDK-Tag using Mouse anti DDDDK-Tag mAb (AE005) at 1:20000 dilution incubated overnight at 4℃.Secondary antibody: HRP-conjugated Goat anti-Mouse IgG (H+L)(AS003) at 1:10000 dilution.Lysates/proteins: 20 μg per lane.Blocking buffer: 3% nonfat dry milk in TBST.Detection: ECL Basic Kit (RM00020).Exposure time: 45s.

Western blot analysis of lysates from wild type (WT) and 293F cells transfected with Mouse anti DDDDK-Tag using Mouse anti DDDDK-Tag mAb (AE005) at 1:20000 dilution incubated overnight at 4℃.Secondary antibody: HRP-conjugated Goat anti-Mouse IgG (H+L)(AS003) at 1:10000 dilution.Lysates/proteins: 20 μg per lane.Blocking buffer: 3% nonfat dry milk in TBST.Detection: ECL Basic Kit (RM00020).Exposure time: 45s.

Western blot analysis of lysates from wild type (WT) and 293F cells transfected with Mouse anti DDDDK-Tag using Mouse anti DDDDK-Tag mAb (AE005) at 1:20000 dilution incubated overnight at 4℃.Secondary antibody: HRP-conjugated Goat anti-Mouse IgG (H+L)(AS003) at 1:10000 dilution.Lysates/proteins: 20 μg per lane.Blocking buffer: 3% nonfat dry milk in TBST.Detection: ECL Basic Kit (RM00020).Exposure time: 45s.

Immunofluorescence analysis of 293T cells using Mouse anti DDDDK-Tag mAb (AE005) at dilution of 1:100 (40x lens). Blue: DAPI for nuclear staining.

Immunofluorescence analysis of 293T-BCAT2-Flag-GFP-C cells using Mouse anti DDDDK-Tag mAb (AE005) at dilution of 1:100 (40x lens). Blue: DAPI for nuclear staining.

Immunofluorescence analysis of 293T-BCAT2-Flag-GFP-N cells using Mouse anti DDDDK-Tag mAb (AE005) at dilution of 1:100 (40x lens). Blue: DAPI for nuclear staining.

Immunoprecipitation of SERPINB1-Flag from 150 µg extracts of 293T cells transfected with a SERPINB1 expression vector containing a single N-terminal DDDDK-Tag was performed using 0.5 µg of Mouse anti DDDDK-Tag mAb (AE005). Mouse IgG isotype control (AC011) was used to precipitate the Control IgG sample. IP samples were eluted with 1X Laemmli Buffer. The Input lane represents 10 % of the total input. Western blot analysis of immunoprecipitates was conducted using Mouse anti DDDDK-Tag mAb (AE005) at a dilution of 1:5000.

Flow cytometry: 1X10^6 CHO cells (negative control,left) and CHO-Claudin18.2-Flag (Transfection,right) cells were intracellularly-stained with Mouse anti DDDDK-Tag mAb (AE005,2 μg/mL,orange line) or Mouse isotype control (2 μg/mL,blue line), followed by PE Goat anti-Mouse pAb staining. Non-fluorescently stained cells were used as blank control (red line).

Flow cytometry: 1X10^6 CHO-Claudin18.2-Flag (Transfection,right) cells were intracellularly-stained with Mouse isotype control (2 μg/mL,left) or Mouse anti DDDDK-Tag mAb (AE005,2 μg/mL,right), followed by PE Goat anti-Mouse pAb staining.

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货号: AE005

促销价：   ￥630
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详细信息

推荐稀释比
  • WB1:20000 - 1:80000
  • IF/ICC1:50 - 1:200
  • IP0.5μg-4μg antibody for 100μg-200μg extracts of whole cells
  • FC (intra)5 μl per 10^6 cells in 100 μl volume
  • ELISARecommended starting concentration is 1 μg/mL. Please optimize the concentration based on your specific assay requirements.
浓度查询

请输入产品标签上的lot号，例如4000000001

反应物种
Species independent
同种型
IgG1,Kappa
实际分子量
37kDa/48kDa/57kDa/70kDa
克隆号
AMC0382
产品形式
Liquid
偶联物
Unconjugated
存储缓冲液
Store at -20℃. Avoid freeze / thaw cycles.
Buffer: PBS with 0.09% Sodium azide,50% glycerol,pH7.3.
阳性样本
293F,293T cells transfected with a SERPINB1
纯化方式
Affinity purification

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用户验证的应用

应用及物种
  • WB (Homo sapiens , G.hirsutum , Phytophthora capsici , Stemphylium eturmiunum , Mus musculus , Rattus norvegicus , Oryctolagus cuniculus , Homo sapiens , Mus musculus , Arabidopsis thaliana , Drosophila melanogaster , Nepenthes , Saccharomyces cerevisiae , Bacillus altitudinis , Capra hircus , Glycine max , Zea mays , Triticum aestivum , Sus scrofa , Other , Oryza sativa , Cyprinus carpio , N. benthamiana , Lycopersicon esculentum , Takifugu rubripes , Escherichia coli , Grapevine , Ctenopharyngodon idellus , Cherax quadricarinatus , Chlorocebus aethiops , D. melanogaste , Danio rerio , Ictalurus punctatus , Cyprinus Carpio , Nicotiana benthamiana , Xenopus laevis , Sus scrofa , Caenorhabditis elegans , Pimephales promelas)
  • IF (Homo sapiens , Bos taurus , Rattus norvegicus , Mus musculus , Escherichia coli , Arabidopsis thaliana , Paralichthys olivaceus , Gallus gallus , D. melanogaste , Other , Danio rerio , Sus scrofa)
  • IP (Homo sapiens , Other , Nepenthes , Mus musculus , Helicoverpa armigera , Nicotiana benthamiana , Drosophila melanogaster , Yeast , Danio rerio , Ictalurus punctatus , Cyprinus carpio , Homo sapiens , Sus scrofa)
  • Co-IP (Stemphylium eturmiunum , Oryza sativa , Arabidopsis thaliana , Homo sapiens , Mus musculus , Other , Danio rerio , Sus scrofa , Xenopus laevis , Caenorhabditis elegans)
  • FC (Homo sapiens)
  • IHC (Homo sapiens , Homo sapiens , Mus musculus)
  • ELISA (Mus musculus , Drosophila melanogaster)
  • ChIP (Mus musculus , Other , Arabidopsis thaliana , Drosophila melanogaster)
  • RIP (Homo sapiens , Drosophila melanogaster)
  • ChIP-Seq (Saccharomyces cerevisiae)
  • Other (Mus musculus)

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文献引用 (249)

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Publishing research using AE005? Please let us know so that we can cite the reference in this datasheet.

背景信息

FLAG-tag, or FLAG octapeptide, or FLAG epitope, is a polypeptide protein tag that can be added to a protein using recombinant DNA technology, having the sequence motif DYKDDDDK. It has been used for studying proteins in living cells and for protein purification by affinity chromatography. It has been used to separate recombinant, overexpressed protein from wild-type protein expressed by the host organism. It can also be used in the isolation of protein complexes with multiple subunits, because its mild purification procedure tends not to disrupt such complexes. It has been used to obtain proteins of sufficient purity and quality to carry out 3D structure determination by x-ray crystallography.A FLAG-tag can be used in many different assays that require recognition by an antibody. If there is no antibody against a given protein, adding a FLAG-tag to a protein allows the protein to be studied with an antibody against the FLAG sequence. Examples are cellular localization studies by immunofluorescence or detection by SDS PAGE protein electrophoresis and Western blotting.

RRID
AB_2770401
别名
DDDDK;DDDDK tag;DDDDK-tag

抗原信息

免疫原
A synthetic peptide corresponding to DDDDK tag.
序列
DYKDDDDK
进行序列比对

FAQ

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ABclonal公司的抗体，可以回收利用几次？

首先，一般抗体不推荐客户回收利用，抗体使用之后缓冲体系已经发生改变，不同客户在回收抗体的保存条件上也会有差异，所以抗体回收使用效果无法保证。另外，ABclonal公司也做过一批抗体回收验证测试，测试结果显示不同抗体可回收次数不同，一般效价越高的抗体，可重复使用的次数越多，客户可根据实验情况来确定。
注：我们将孵育完毕后剩余的抗体回收到离心管中置于4℃保存，效价高的抗体可至少保存1周，至少重复利用3次。

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ABclonal作为国产抗体品牌的优势？

武汉爱博泰克生物（ABclonal）科技有限公司是国产品牌，她成立于2011年，公司依托ABclonal美国波士顿抗体与蛋白研发中心、中国光谷生物城(武汉)抗体生产基地以及上海张江分子酶研发中心，凝聚了十余位来自哈佛大学、麻省理工、复旦大学、上海交大、中科院生化细胞所和武汉大学的全球知名分子和免疫学方面博士，组成我们的科学家团队，通过聚焦抗体与酶核心技术，致力于打破国际技术的垄断，将公司打造成为科研工具和诊断原料的国内领导品牌，乃至弯道超越国际巨头。 我们拥有包括兔多克隆抗体、小鼠单克隆抗体、兔单克隆抗体的生产研发平台，同时也有包括WB,IHC,IF,IP,CHIP在内的检测平台，我们对每一支自产的抗体进行了严格的检测。当然，我们部分直销地区也可以帮客户代购进口品牌的产品。同时也有抗体定制服务。ABclonal抗体优势：1，严自检，保质量；2产品多，指标全；3，价格低，货期短。注：ABclonal抗体价格体系详情见附件

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ABclonal抗体成分？

ABclonal抗体成分在平时工作当中，常会有客户咨询我们的抗体用的什么buffer进行保存，一般来说，我们的buffer的成分是：PBS含0.03%的proclin300、0.05%牛血清白蛋白、50%甘油；也有一些是PBS含0.03%的proclin300，50%甘油。防腐剂 Proclin 300活性成分主要是2-甲基-4-异噻唑啉-3-酮（MCI）和5-氯-2-甲基-4-异噻唑啉-3-酮（CMCI）。ProClin生物灭活剂能够迅速穿透细胞膜，抑制对细胞呼吸至关重要的特定酶，因此一接触微生物有机体就会立即抑制细胞活性。ProClin的多个特定毒性位点可以防止微生物产生高水平的耐药性。

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实验方案

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