DYKDDDDK Tag Rabbit mAb

Catalog No.: F3506

Application: Reactivity: All

Usage Information

Dilution
WB1:1000 IP1:50 IHC1:400 - 1:1600 IF1:400 - 1:1600 FCM1:800 - 1:3200 CHIP1:50

Application
WB, IP, IHC, IF, F, ChIP

Source
Rabbit

Reactivity
All

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Datasheet & SDS

Biological Description

Specificity
DYKDDDDK Tag Rabbit mAb detects exogenously expressed DYKDDDDK proteins in cells. The antibody recognizes the DYKDDDDK peptide, which is the same epitope recognized by Sigma-Aldrich Anti-FLAG M2 antibody, fused to either the amino-terminus or carboxy-terminus of the target protein.

Synonym(s)
DDK; FLAG; Flag tag; tag

Clone
C19M9

Background
The DYKDDDDK tag, commonly known as the FLAG tag, is an eight–amino acid epitope tag (Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys), widely used in molecular biology for protein purification, detection, and expression studies. This tag is genetically fused to either the N-terminal or C-terminal of a target protein using recombinant DNA technology, and its small size minimizes interference with the protein's function. The FLAG tag allows highly specific purification via affinity chromatography by binding to monoclonal antibodies, such as anti-FLAG M2 (position-insensitive) or M1 (N-terminal specific and calcium-dependent). Proteins fused with FLAG can be eluted from the matrix using a low pH buffer or excess FLAG peptide, and the tag can be removed enzymatically at a protease cleavage site (e.g., enterokinase). Tandem FLAG tags (e.g., 3xFLAG or modified sequences like DYKDHDG) are used for enhanced binding and detection. The FLAG tag is extensively applied in assays such as western blotting, ELISA, immunoprecipitation, flow cytometry, immunofluorescence, and SDS-PAGE, particularly in mammalian expression systems, and can also be combined with other epitope tags (e.g., HA, His, Myc) for multi-functional studies.

Background

The DYKDDDDK tag, commonly known as the FLAG tag, is an eight–amino acid epitope tag (Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys), widely used in molecular biology for protein purification, detection, and expression studies. This tag is genetically fused to either the N-terminal or C-terminal of a target protein using recombinant DNA technology, and its small size minimizes interference with the protein's function. The FLAG tag allows highly specific purification via affinity chromatography by binding to monoclonal antibodies, such as anti-FLAG M2 (position-insensitive) or M1 (N-terminal specific and calcium-dependent). Proteins fused with FLAG can be eluted from the matrix using a low pH buffer or excess FLAG peptide, and the tag can be removed enzymatically at a protease cleavage site (e.g., enterokinase). Tandem FLAG tags (e.g., 3xFLAG or modified sequences like DYKDHDG) are used for enhanced binding and detection. The FLAG tag is extensively applied in assays such as western blotting, ELISA, immunoprecipitation, flow cytometry, immunofluorescence, and SDS-PAGE, particularly in mammalian expression systems, and can also be combined with other epitope tags (e.g., HA, His, Myc) for multi-functional studies.

References
[1] Brizzard BL, et al. Biotechniques. 1994 Apr;16(4):730-5.

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

Tel: +1-832-582-8158 Ext:3
If you have any other enquiries, please leave a message.

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%