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DDDDK-Tag Rabbit mAb (AE063)

Publications (72) Datasheet

Tested applications:WBIHC-PIF/ICCIPChIPChIP-seqRIPFCFC(Intra)ELISAMeDIPNucleotide ArrayDBFACSCoIPCUT&TagmeRIPInhibitionReactivity:Species independent

ABclonal:DDDDK-Tag Rabbit mAb
ABclonal:Western blot - DDDDK-Tag Rabbit mAb (AE063)

Western blot analysis of lysates from from wild type (WT) and 293T cells transfected with GSK3B-C using DDDDK-Tag Rabbit mAb (AE063) at 1:5000 dilution.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 30s.

ABclonal:Immunofluorescence - DDDDK-Tag Rabbit mAb (AE063)

Immunofluorescence analysis of GFP-DDDDK transgenic HeLa cells using DDDDK-Tag Rabbit mAb (AE063). Green: GFP expression. Blue: DAPI for nuclear staining. 

ABclonal:Immunofluorescence - DDDDK-Tag Rabbit mAb (AE063)

Immunofluorescence analysis of 293T-Flag-C and 293T-Flag-N and 293T cells using DDDDK-Tag Rabbit mAb (AE063) at dilution of 1:100 (40x lens). Secondary antibody: Cy3-conjugated Goat anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.

ABclonal:Immunoprecipitation - DDDDK-Tag Rabbit mAb (AE063)

Immunoprecipitation of over-expressed DDDDK-tagged protein in 293T cells incubated using DDDDK-tag antibody (AE063), Secondary antibody: HRP-conjugated AffiniPure Mouse Anti-Rabbit IgG Light Chain (AS061). A mock served as negative control and over-expressed 293T cell lysate served as positive control.

ABclonal:Immunoprecipitation - DDDDK-Tag Rabbit mAb (AE063)

Immunoprecipitation of IRF1-Flag from 600 µg extracts of 293T cells transfected with a IRF1 expression vector containing a single N-terminal Flag-Tag was performed using 3 µg of DDDDK-Tag Rabbit mAb (AE063). Rabbit IgG isotype control (AC042) was used to precipitate the Control IgG sample. The IP sample was eluted with 1X reducing Laemmli Buffer. The Input lane represents 10% of the total input. Western blot analysis of immunoprecipitates was conducted using DDDDK-Tag Rabbit mAb (AE063) at a dilution of 1:1000.

Overview

Product nameDDDDK-Tag Rabbit mAb
Catalog No.AE063
Host speciesRabbit
Purification methodAffinity purification
IsotypeIgG
CloneNo.ARC5111-02
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.
ImmunogenA synthetic peptide corresponding to DDDDK tag.
SequenceDYKDDDDK
Gene ID
Swiss Prot
SynonymsDDDDK; DDDDK tag; DDDDK-tag; DDDDK-Tag
Calculated MW
Observed MWRefer to Figures
ReactivitySpecies independent
Tested applicationsWBIHC-PIF/ICCIPChIPChIP-seqRIPFCFC(Intra)ELISAMeDIPNucleotide ArrayDBFACSCoIPCUT&TagmeRIPInhibition
Recommended dilution
  • WB 1:2000 - 1:6000
  • IF/ICC 1:50 - 1:200
  • IP 0.5μg-4μg antibody for 200μg-400μg extracts of whole cells
  • ELISA Recommended starting concentration is 1 μg/mL. Please optimize the concentration based on your specific assay requirements.
Storage bufferStore at -20℃. Avoid freeze / thaw cycles.
Buffer: PBS with 0.05% proclin300, 0.05% BSA, 50% glycerol, pH7.3.
Key applicationWestern blotting    Immunofluorescence    Immunoprecipitation    
Positive samples293T
Cellular location
Customer validation

WB(Oreochroms mossambcus, Homo sapiens, Mus musculus, Arabidopsis thaliana, Bombyx mori Linnaeus, Mus musculus, Other, Triticum aestivum, Gallus gallus)

IHC(Mus musculus, Homo sapiens)

IP(Arabidopsis thaliana)

RIP(Homo sapiens)

IP(Homo sapiens, Mus musculus, Other)

FC(Homo sapiens)

IP(Homo sapiens, Mus musculus, Other)

Co-IP(Homo sapiens)

IF(Other)

ELISA(Other)

WB(Mus musculus, Other)

ChIP(Arabidopsis thaliana, Other)

IHC(Mus musculus)

IF(Mus musculus, Homo sapiens)

Co-IP(Glycine max, Homo sapiens)

PCR(Other)

RT-PCR(Mus musculus)

RNA-Seq(Mus musculus)

Documents

Certificate of Compliance

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Lot number

    ABclonal:Western blot - DDDDK-Tag Rabbit mAb (AE063)}

    Western blot - DDDDK-Tag Rabbit mAb (AE063)

    Western blot analysis of lysates from from wild type (WT) and 293T cells transfected with GSK3B-C using DDDDK-Tag Rabbit mAb (AE063) at 1:5000 dilution.
    Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
    Lysates/proteins: 25μg per lane.
    Blocking buffer: 3% nonfat dry milk in TBST.
    Detection: ECL Basic Kit (RM00020).
    Exposure time: 30s.
    ABclonal:Immunofluorescence - DDDDK-Tag Rabbit mAb (AE063)}

    Immunofluorescence - DDDDK-Tag Rabbit mAb (AE063)

    Immunofluorescence analysis of GFP-DDDDK transgenic HeLa cells using DDDDK-Tag Rabbit mAb (AE063). Green: GFP expression. Blue: DAPI for nuclear staining. 
    ABclonal:Immunofluorescence - DDDDK-Tag Rabbit mAb (AE063)}

    Immunofluorescence - DDDDK-Tag Rabbit mAb (AE063)

    Immunofluorescence analysis of 293T-Flag-C and 293T-Flag-N and 293T cells using DDDDK-Tag Rabbit mAb (AE063) at dilution of 1:100 (40x lens). Secondary antibody: Cy3-conjugated Goat anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.
    ABclonal:Immunoprecipitation - DDDDK-Tag Rabbit mAb (AE063)}

    Immunoprecipitation - DDDDK-Tag Rabbit mAb (AE063)

    Immunoprecipitation of over-expressed DDDDK-tagged protein in 293T cells incubated using DDDDK-tag antibody (AE063), Secondary antibody: HRP-conjugated AffiniPure Mouse Anti-Rabbit IgG Light Chain (AS061). A mock served as negative control and over-expressed 293T cell lysate served as positive control.
    ABclonal:Immunoprecipitation - DDDDK-Tag Rabbit mAb (AE063)}

    Immunoprecipitation - DDDDK-Tag Rabbit mAb (AE063)

    Immunoprecipitation of IRF1-Flag from 600 µg extracts of 293T cells transfected with a IRF1 expression vector containing a single N-terminal Flag-Tag was performed using 3 µg of DDDDK-Tag Rabbit mAb (AE063). Rabbit IgG isotype control (AC042) was used to precipitate the Control IgG sample. The IP sample was eluted with 1X reducing Laemmli Buffer. The Input lane represents 10% of the total input. Western blot analysis of immunoprecipitates was conducted using DDDDK-Tag Rabbit mAb (AE063) at a dilution of 1:1000.

    * For research use only. Not for therapeutic or diagnostic purposes.

    Publishing research using AE063? Please let us know so that we can cite the reference in this datasheet.