Cell Signaling Technology

Product Pathways - Tag Antibodies

DYKDDDDK Tag (D6W5B) Rabbit mAb (Binds to same epitope as Sigma's Anti-FLAG® M2 Antibody) #14793

DDK   FLAG   Flag tag   tag  

No. Size Price
14793S 100 µl ( 10 western blots ) ¥3,100.00 现货查询 购买询价
14793 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 All Species Expected, Transfected Only Rabbit IgG
IP 1:50
IHC-P 1:800
F 1:1600
IF-IC 1:800
ChIP 1:50

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), F=Flow Cytometry, IF-IC=Immunofluorescence (Immunocytochemistry), ChIP=Chromatin IP,

Specificity / Sensitivity

DYKDDDDK Tag (D6W5B) Rabbit mAb detects exogenously expressed DYKDDDDK proteins in cells. The antibody recognizes the DYKDDDDK peptide, which is the same epitope recognized by Sigma's Anti-FLAG® antibodies, fused to either the amino-terminus or carboxy-terminus of the target protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic DYKDDDDK peptide.

Flow Cytometry

Flow Cytometry

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.

IF-IC

IF-IC

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 Blotting

Western Blotting

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.

IHC-P (paraffin)

IHC-P (paraffin)

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

Chromatin IP

Chromatin IP

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.

IP

IP

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.

Background

Epitope tags are useful for the labeling and detection of proteins using immunoblotting, immunoprecipitation, and immunostaining techniques. Because of their small size, they are unlikely to affect the biochemical properties of tagged proteins.

The DYKDDDDK peptide has been used extensively as a general epitope tag in expression vectors. This peptide can be expressed and detected with the protein of interest as an amino-terminal or carboxy-terminal fusion (1).

  1. Brizzard, B.L. et al. (1994) Biotechniques 16, 730-5.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Protocols

Companion Products


For Research Use Only. Not For Use In Diagnostic Procedures.

Anti-FLAG is a registered trademark of Sigma-Aldrich Biotechnology.

DRAQ5 is a registered trademark of Biostatus Limited.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

XP is a registered trademark of Cell Signaling Technology, Inc.

SignalStain is a trademark of Cell Signaling Technology, Inc.

SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.

Alexa Fluor is a registered trademark of Life Technologies Corporation.

Tween is a registered trademark of ICI Americas, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

用户评论 --- 共 0

该产品暂无评论!

我要参与评论 :

如要参与评论请先登录网站

还没有网站账户?去注册一下吧

Products

 

Applications