DYKDDDDK tag Recombinant antibody (Binds to FLAG® tag epitope)

DYKDDDDK tag Uni-rAb^TM Recombinant Antibody for WB, IF/ICC, FC (Intra), IP, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

recombinant protein

Applications

WB, IF/ICC, FC (Intra), IP, ELISA

Conjugate

Unconjugated

CloneNo.

240568A11

Cat no : 80801-2-RR

Synonyms

3Flag, Flag, DDDDKtag, DDDDK tag, DDDDK

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Validation Data Gallery

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  WB analysis of Transfected HEK-293T using 80801-2-RR

  TDP-43-flag Transfected HEK-293T cells were subjected to SDS PAGE followed by western blot with 80801-2-RR (Ag2329 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  IP experiment of Transfected HEK-293 using 80801-2-RR

  IP result of anti-DYKDDDDK tag (IP:80801-2-RR, 4ug; Detection:80801-2-RR 1:3000) with Transfected HEK-293 cells lysate 400 ug.

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  IF Staining of Transfected HEK-293T using 80801-2-RR

  Immunofluorescent analysis of (4% PFA) fixed Transfected HEK-293T cells using DYKDDDDK tag antibody (80801-2-RR, Clone: 240568A11 ) at dilution of 1:250 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2).

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  FC experiment of HEK-293T transfected with TDP-43-flag using 80801-2-RR

  1x10^6 HEK-293T transfected with TDP-43-flag were intracellularly stained with 0.4 ug DYKDDDDK tag Recombinant antibody (Binds to FLAG® tag epitope) (80801-2-RR, Clone:240568A11) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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  Affinity and Kinetic Characterization of 80801-2-RR

  Biolayer interferometry (BLl) kinetic assays of 80801-2-RR against DYKDDDDK tag were performed. The affinity constant is 15.3 pM.

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Tested Applications

• Positive WB detected in: TDP-43-flag Transfected HEK-293T cells
• Positive IP detected in: Transfected HEK-293 cells
• Positive IF/ICC detected in: Transfected HEK-293T cells
• Positive FC (Intra) detected in: HEK-293T transfected with TDP-43-flag

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunofluorescence (IF)/ICC: IF/ICC : 1:125-1:500
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

The immunogen of 80801-2-RR is DYKDDDDK tag Fusion Protein expressed in E. coli.

• Tested Reactivity: recombinant protein
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: DYKDDDDK tag fusion protein Ag2329
• Full Name: DYKDDDDK tag
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purfication
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Protein tags are protein or peptide sequences located either on the C- or N- terminal of the target protein, which facilitates one or several of the following characteristics: solubility, detection, purification, localization and expression. The DYKDDDDK(FLAG) peptide has been used extensively as a general tag in expression vectors. This peptide can be expressed and detected with the protein of interest as an amino-terminal or carboxy-terminal fusion. N-terminal DDDDK vectors provide an Ek cleavage site for removal of the fusion tag. The DDDDK peptide is likely to be located on the surface of a fusion protein because of its hydrophilic nature. As a result, the DDDDK peptide is more likely to be accessible to antibodies. A DDDDK-tag can be used in many different assays that require recognition by an antibody, such as western blotting, immunocytochemistry, immunoprecipitation, flow cytometry, protein purification, and in the study of protein-protein interactions, cell ultrastructure, and protein localization and so on. This antibody is a rabbit polyclonal antibody raised against 3xFlag (3xDYKDDDDKT) sequence and recognizes the (1x) and (3x)DYKDDDDK peptide and detects DDDDK-tagged proteins. Anti-FLAG is a registered trademark of Sigma-Aldrich Biotechnology.