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DYKDDDDK Tag (D6W5B) Rabbit mAb (Binds to same epitope as Sigma-Aldrich Anti-FLAG M2 antibody)

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DYKDDDDK Tag (D6W5B) Rabbit mAb (Binds to same epitope as Sigma-Aldrich Anti-FLAG M2 antibody)

品牌： CST

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参数

反应种属：

All

来源宿主：

Rabbit

产品介绍

产品信息

抗原名称

DYKDDDDK Tag

来源纯化

使用合成的 DYKDDDDK 肽，对动物进行免疫接种来产生单克隆抗体。

宿主

Rabbit

简单描述

Monoclonal Antibody for studying Flag-Tag. Cited in 1073 publications. Validated for WB, IP, IHC, IF, F, ChIP. Available in 2 sizes. Highly specific and rigorously validated in-house, DYKDDDDK Tag (D6W5B) Rabbit Monoclonal Antibody (Binds to same epitope as Sigma-Aldrich Anti-FLAG M2 antibody) (CST #14793) is ready to ship.

商品描述

Product Usage Information

For optimal ChIP results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

Application	Dilution
Western Blotting	1:1000
Immunoprecipitation	1:50
Immunohistochemistry (Paraffin)	1:400 - 1:1600
Immunofluorescence (Immunocytochemistry)	1:400 - 1:1600
Flow Cytometry (Fixed/Permeabilized)	1:800 - 1:3200
Chromatin IP	1:50

应用

反应种属

All

目标/特异性

Specificity/Sensitivity

DYKDDDDK Tag (D6W5B) Rabbit mAb 可检测细胞内 DYKDDDDK 蛋白的外生性表达。这种抗体识别与所靶向蛋白质的氨基末端或羧基末端融合的 DYKDDDDK 肽，该肽是由 Sigma-Aldrich 抗 FLAG M2 抗体识别的相同表位。

物种反应性：

预期的所有物种

敏感性

Transfected Only

背景

在使用免疫印迹法、免疫沉淀法和免疫染色技术对蛋白进行标记和检测时，表位标签十分有用。由于分子量小，它们不太可能对标记蛋白的生化特性造成影响。DYKDDDDK 肽在表达载体中广泛用作附加标签。该肽段可以与目标蛋白融合，作为氨基末端或羧基末端表达，并能够被检测 (1)。 1.Brizzard, B.L. et al. (1994) Biotechniques 16, 730-5.

翻译后修饰

Unmodified

制备和贮存

保存方式

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.For a carrier free (BSA and azide free) version of this product see product #70586.

参考图片

Confocal immunofluorescent analysis of 293T cells transfected with a GFP-DYKDDDDK-Tag (green) using DYKDDDDK Tag (D6W5B)Rabbit mAb (red). Blue pseudocolor = DRAQ5^® #4084 (fluorescent DNA dye).

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with DYKDDDDK-GFP (N-terminal DDK-Tag; +), GFP-DYKDDDDK (C-terminal DDK-Tag; +), human CASQ1-DYKDDDDK (C-terminal DDK-Tag; +), or human FoxG1-DYKDDDDK (C-terminal DDK-Tag; +) as indicated, using DYKDDDDK Tag (D6W5B)Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded HeLa cell pellets, control (left) or DYKDDDDK Tag-transfected (right), using DYKDDDDK Tag (D6W5B) Rabbit mAb.

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10⁶ 293T cells, untransfected (left) or transfected (right) with a DYKDDDDK-tagged SSRP1 construct and 10 µl of DYKDDDDK (D6W5B)Rabbit mAb, 10 µl of SSRP1 (E1Y8D) Rabbit mAb #13421, or 2 µl of Normal Rabbit IgG #2729, using SimpleChIP^® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP^® Human c-Fos Exon 3 Primers #12010 and SimpleChIP^® Human EGR1 Intron 3 Primers #11953. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

Immunoprecipitation of FoxG1-DYKDDDDK tag protein from 293T transfected cell extracts using Rabbit (DA1E) mAb IgG XP^® Isotype Control #3900 (lane 2) or DYKDDDDK (D6W5B)Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using DYKDDDDK (D6W5B)Rabbit mAb.

Flow cytometric analysis of 293 cells, untransfected (blue) or transfected with DYKDDDDK-Tag (green), using DYKDDDDK Tag (D6W5B)Rabbit mAb. Anti-Rabbit IgG (H+L), F(ab')₂ Fragment (Alexa Fluor^® 488 Conjugate) #4412 was used as a secondary antibody.

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货号：

14793S

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