Product Type > Antibodies > Tag and Loading Control Antibodies

Rabbit anti DDDDK-Tag pAb (AE004)

Publications (27) Datasheet SDS COA

Tested applications: WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra) ELISA MeDIP Nucleotide Array DB FACS CoIP FCM CUT&Tag meRIP Inhibition

Reactivity:Species independent

ABclonal:Western blot - Rabbit anti DDDDK-Tag pAb (AE004)

Western blot analysis of lysates from wild type (WT) and 293T cells transfected with DDDDK-Tag using Rabbit anti DDDDK-Tag pAb(AE004) at 1:10000 dilution. 
Secondary antibody:(AS014 ) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection:ECL Basic Kit (RM00020).
Exposuretime: 1s.

ABclonal:Immunofluorescence - Rabbit anti DDDDK-Tag pAb (AE004)

Immunofluorescence analysis of 293T cells transfected with Flag-c(mCherry) and 293T cells transfected with Flag-N(mCherry) use Rabbit anti DDDDK-Tag pAb (AE004) at dilution of 1:50 (40x lens). Secondary antibody: Cy3 Goat Anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.

ABclonal:Immunofluorescence - Rabbit anti DDDDK-Tag pAb (AE004)

Immunofluorescence analysis of 293T cells transfected with Flag-N(mCherry) and untreated 293T cells use Rabbit anti DDDDK-Tag pAb (AE004) at dilution of 1:100 (40x lens). Secondary antibody: Cy3 Goat Anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.

ABclonal:Immunoprecipitation - Rabbit anti DDDDK-Tag pAb (AE004)

Immunoprecipitation analysis of 300 μg extracts of 293T-SERPINB1-Flag-GFP cells using 3 μg DDDDK-Tag pAb (AE004). Western blot was performed from the immunoprecipitate using DDDDK-Tag pAb (AE004) at a dilition of 1:50000.

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Overview

Product name Rabbit anti DDDDK-Tag pAb
Catalog No. AE004
Host species Rabbit
Purification method Affinity purification
Isotype IgG

Background

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.

Immunogen information

Immunogen A synthetic peptide corresponding to DDDDK tag.
Sequence DYKDDDDK
Gene ID
Swiss prot
Synonyms DDDDK; DDDDK tag; DDDDK-tag
Calculated MW
Observed MW 48kDa

Applications

Reactivity Species independent
Tested applications WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra) ELISA MeDIP Nucleotide Array DB FACS CoIP FCM CUT&Tag meRIP Inhibition
Recommended dilution
  • WB 1:2000 - 1:10000
  • IF/ICC 1:50 - 1:200
  • IP 0.5μg-4μg antibody for 200μg-400μg extracts of whole cells
Storage buffer Store at -20℃. Avoid freeze / thaw cycles.
Buffer: PBS with 0.05% proclin300, 50% glycerol, pH7.3.
Application key Western blotting    Immunofluorescence    Immunoprecipitation    
Positive samples 293T
Cellular location
Customer validation

WB (Homo sapiens, Mus musculus, Mesocricetus auratus, Sus scrofa, Rattus norvegicus, Haloferax volcanii)

IP (Oryza sativa, Homo sapiens, Danio rerio, Xanthonycticebus pygmaeus, Mus musculus, Haloferax volcanii)

Co-IP (Homo sapiens)

IHC (Mus musculus)

IF (Homo sapiens, Sus scrofa)

ChIP (Homo sapiens)

Documents

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

Lot number

Rabbit anti DDDDK-Tag pAb images

ABclonal:Western blot - Rabbit anti DDDDK-Tag pAb (AE004)}

Western blot - Rabbit anti DDDDK-Tag pAb (AE004)

Western blot analysis of lysates from wild type (WT) and 293T cells transfected with DDDDK-Tag using Rabbit anti DDDDK-Tag pAb(AE004) at 1:10000 dilution. 
Secondary antibody:(AS014 ) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection:ECL Basic Kit (RM00020).
Exposuretime: 1s.
ABclonal:Immunofluorescence - Rabbit anti DDDDK-Tag pAb (AE004)}

Immunofluorescence - Rabbit anti DDDDK-Tag pAb (AE004)

Immunofluorescence analysis of 293T cells transfected with Flag-c(mCherry) and 293T cells transfected with Flag-N(mCherry) use Rabbit anti DDDDK-Tag pAb (AE004) at dilution of 1:50 (40x lens). Secondary antibody: Cy3 Goat Anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.
ABclonal:Immunofluorescence - Rabbit anti DDDDK-Tag pAb (AE004)}

Immunofluorescence - Rabbit anti DDDDK-Tag pAb (AE004)

Immunofluorescence analysis of 293T cells transfected with Flag-N(mCherry) and untreated 293T cells use Rabbit anti DDDDK-Tag pAb (AE004) at dilution of 1:100 (40x lens). Secondary antibody: Cy3 Goat Anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.
ABclonal:Immunoprecipitation - Rabbit anti DDDDK-Tag pAb (AE004)}

Immunoprecipitation - Rabbit anti DDDDK-Tag pAb (AE004)

Immunoprecipitation analysis of 300 μg extracts of 293T-SERPINB1-Flag-GFP cells using 3 μg DDDDK-Tag pAb (AE004). Western blot was performed from the immunoprecipitate using DDDDK-Tag pAb (AE004) at a dilition of 1:50000.

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Related Targets

Based on existing publications, the following genes are closely related to research on DDDDK. (Numbers indicate number of published articles where both targets appear.)

1/

Related Research Area

Based on existing publications, the following research topics are related to DDDDK. (Distance between topics and target gene indicate popularity.) DDDDK

* Data provided by citexs.com, for reference only.

Publishing research using AE004? Please let us know so that we can cite the reference in this datasheet.

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

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