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Mouse anti DDDDK-Tag mAb (AE005)

Publications (245) Datasheet

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

ABclonal:Western blot - Mouse anti DDDDK-Tag mAb (AE005)

Western blot analysis of lysates from wild type (WT) and 293F cells transfected with Mouse anti DDDDK-Tag using Mouse anti DDDDK-Tag mAb (AE005) at 1:20000 dilution incubated overnight at 4℃.
Secondary antibody: HRP-conjugated Goat anti-Mouse IgG (H+L)(AS003) at 1:10000 dilution.
Lysates/proteins: 20 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020)
.Exposure time: 45s.

ABclonal:Western blot - Mouse anti DDDDK-Tag mAb (AE005)

Western blot analysis of lysates from wild type (WT) and 293F cells transfected with Mouse anti DDDDK-Tag using Mouse anti DDDDK-Tag mAb (AE005) at 1:20000 dilution incubated overnight at 4℃.
Secondary antibody: HRP-conjugated Goat anti-Mouse IgG (H+L)(AS003) at 1:10000 dilution.
Lysates/proteins: 20 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020)
.Exposure time: 45s.

ABclonal:Western blot - Mouse anti DDDDK-Tag mAb (AE005)

Western blot analysis of lysates from wild type (WT) and 293F cells transfected with Mouse anti DDDDK-Tag using Mouse anti DDDDK-Tag mAb (AE005) at 1:20000 dilution incubated overnight at 4℃.
Secondary antibody: HRP-conjugated Goat anti-Mouse IgG (H+L)(AS003) at 1:10000 dilution.
Lysates/proteins: 20 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020)
.Exposure time: 45s.

ABclonal:Immunofluorescence - Mouse anti DDDDK-Tag mAb (AE005)

Immunofluorescence analysis of 293T cells using Mouse anti DDDDK-Tag mAb (AE005) at dilution of 1:100 (40x lens). Blue: DAPI for nuclear staining.

ABclonal:Immunofluorescence - Mouse anti DDDDK-Tag mAb (AE005)

Immunofluorescence analysis of 293T-BCAT2-Flag-GFP-C cells using Mouse anti DDDDK-Tag mAb (AE005) at dilution of 1:100 (40x lens). Blue: DAPI for nuclear staining.

ABclonal:Immunofluorescence - Mouse anti DDDDK-Tag mAb (AE005)

Immunofluorescence analysis of 293T-BCAT2-Flag-GFP-N cells using Mouse anti DDDDK-Tag mAb (AE005) at dilution of 1:100 (40x lens). Blue: DAPI for nuclear staining.

ABclonal:Immunoprecipitation - Mouse anti DDDDK-Tag mAb (AE005)

Immunoprecipitation of SERPINB1-Flag from 150 µg extracts of 293T cells transfected with a SERPINB1 expression vector containing a single N-terminal DDDDK-Tag was performed using 0.5 µg of Mouse anti DDDDK-Tag mAb (AE005). Mouse IgG isotype control (AC011) was used to precipitate the Control IgG sample. IP samples were eluted with 1X Laemmli Buffer. The Input lane represents 10 % of the total input. Western blot analysis of immunoprecipitates was conducted using Mouse anti DDDDK-Tag mAb (AE005) at a dilution of 1:5000.

ABclonal:Flow CytoMetry - Mouse anti DDDDK-Tag mAb (AE005)

Flow cytometry: 1X10^6 CHO cells (negative control, left) and CHO-Claudin18.2-Flag (Transfection, right) cells were intracellularly-stained with Mouse anti DDDDK-Tag mAb (AE005, 2 μg/mL, orange line) or Mouse isotype control (2 μg/mL, blue line), followed by PE Goat anti-Mouse pAb staining. Non-fluorescently stained cells were used as blank control (red line).

ABclonal:Flow CytoMetry - Mouse anti DDDDK-Tag mAb (AE005)

Flow cytometry: 1X10^6 CHO-Claudin18.2-Flag (Transfection, right) cells were intracellularly-stained with Mouse isotype control (2 μg/mL, left) or Mouse anti DDDDK-Tag mAb (AE005, 2 μg/mL, right), followed by PE Goat anti-Mouse pAb staining.

Overview

Product nameMouse anti DDDDK-Tag mAb
Catalog No.AE005
Host speciesMouse
Purification methodAffinity purification
IsotypeIgG1, Kappa
CloneNo.AMC0382
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
Calculated MW
Observed MW37kDa/48kDa/57kDa/70kDa
ReactivitySpecies independent
Tested applicationsWBIHC-PIF/ICCIPChIPChIP-seqRIPFCFC(Intra)ELISAMeDIPNucleotide ArrayDBFACSCoIPCUT&TagmeRIPInhibition
Recommended dilution
  • WB 1:20000 - 1:80000
  • IF/ICC 1:50 - 1:200
  • IP 0.5μg-4μg antibody for 100μg-200μg extracts of whole cells
  • FC (intra) 5 μl per 10^6 cells in 100 μl volume
  • 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 0.09% Sodium azide, 50% glycerol, pH7.3.
Key applicationWestern blotting    Immunofluorescence    Immunoprecipitation    
Positive samples293F, 293T cells transfected with a SERPINB1
Cellular location
Customer validation

WB(Homo sapiens, Oryza sativa, Arabidopsis thaliana, Mus musculus, Sus scrofa, Drosophila melanogaster, Ctenopharyngodon idellus, Escherichia coli, Cyprinus carpio, Grapevine, Nicotiana benthamiana, Danio rerio, Oryctolagus cuniculus, Lycopersicon esculentum, Triticum aestivum, Bacillus altitudinis, Mus musculus, Stemphylium eturmiunum, Capra hircus, Xenopus laevis, Rattus norvegicus, Saccharomyces cerevisiae, D. melanogaste, Cherax quadricarinatus, Sus scrofa, Zea mays, Ictalurus punctatus, , Nepenthes, Caenorhabditis elegans, Other, Glycine max, Takifugu rubripes, Chlorocebus aethiops)

IP(Homo sapiens,Yeast, Danio rerio, Nicotiana benthamiana, Mus musculus, Helicoverpa armigera, Drosophila melanogaster, Homo sapiens, Other, Cyprinus carpio)

IF(Homo sapiens, Bos taurus, Rattus norvegicus, Mus musculus, Escherichia coli, Danio rerio, Paralichthys olivaceus, Arabidopsis thaliana)

Co-IP(Homo sapiens, Other, Oryza sativa, Sus scrofa, Arabidopsis thaliana, Mus musculus)

Co-IP(Danio rerio )

IP(Homo sapiens, Danio rerio, Mus musculus, Sus scrofa, Ictalurus punctatus, Nepenthes)

IF(Homo sapiens, Sus scrofa, Rattus norvegicus, D. melanogaste)

Co-IP(Homo sapiens, Stemphylium eturmiunum, Xenopus laevis, Caenorhabditis elegans)

IHC(Homo sapiens, Mus musculus, )

RIP(Drosophila melanogaster)

ChIP(Drosophila melanogaster, Mus musculus)

WB(Homo sapiens, Mus musculus, Saccharomyces cerevisiae, Cyprinus carpio, Other, Arabidopsis thaliana)

ChIP(Other, Arabidopsis thaliana)

IF(Homo sapiens, Other)

IP(Homo sapiens)

RIP(Homo sapiens)

ELISA(Mus musculus)

ChIP-Seq(Saccharomyces cerevisiae)

ELISA(Drosophila melanogaster)

FC(Homo sapiens)

Other(Mus musculus)

RIP(Mus musculus)

Documents

Certificate of Compliance

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Lot number

    ABclonal:Western blot - Mouse anti DDDDK-Tag mAb (AE005)}

    Western blot - Mouse anti DDDDK-Tag mAb (AE005)

    Western blot analysis of lysates from wild type (WT) and 293F cells transfected with Mouse anti DDDDK-Tag using Mouse anti DDDDK-Tag mAb (AE005) at 1:20000 dilution incubated overnight at 4℃.
    Secondary antibody: HRP-conjugated Goat anti-Mouse IgG (H+L)(AS003) at 1:10000 dilution.
    Lysates/proteins: 20 μg per lane.
    Blocking buffer: 3% nonfat dry milk in TBST.
    Detection: ECL Basic Kit (RM00020)
    .Exposure time: 45s.
    ABclonal:Western blot - Mouse anti DDDDK-Tag mAb (AE005)}

    Western blot - Mouse anti DDDDK-Tag mAb (AE005)

    Western blot analysis of lysates from wild type (WT) and 293F cells transfected with Mouse anti DDDDK-Tag using Mouse anti DDDDK-Tag mAb (AE005) at 1:20000 dilution incubated overnight at 4℃.
    Secondary antibody: HRP-conjugated Goat anti-Mouse IgG (H+L)(AS003) at 1:10000 dilution.
    Lysates/proteins: 20 μg per lane.
    Blocking buffer: 3% nonfat dry milk in TBST.
    Detection: ECL Basic Kit (RM00020)
    .Exposure time: 45s.
    ABclonal:Western blot - Mouse anti DDDDK-Tag mAb (AE005)}

    Western blot - Mouse anti DDDDK-Tag mAb (AE005)

    Western blot analysis of lysates from wild type (WT) and 293F cells transfected with Mouse anti DDDDK-Tag using Mouse anti DDDDK-Tag mAb (AE005) at 1:20000 dilution incubated overnight at 4℃.
    Secondary antibody: HRP-conjugated Goat anti-Mouse IgG (H+L)(AS003) at 1:10000 dilution.
    Lysates/proteins: 20 μg per lane.
    Blocking buffer: 3% nonfat dry milk in TBST.
    Detection: ECL Basic Kit (RM00020)
    .Exposure time: 45s.
    ABclonal:Immunofluorescence - Mouse anti DDDDK-Tag mAb (AE005)}

    Immunofluorescence - Mouse anti DDDDK-Tag mAb (AE005)

    Immunofluorescence analysis of 293T cells using Mouse anti DDDDK-Tag mAb (AE005) at dilution of 1:100 (40x lens). Blue: DAPI for nuclear staining.
    ABclonal:Immunofluorescence - Mouse anti DDDDK-Tag mAb (AE005)}

    Immunofluorescence - Mouse anti DDDDK-Tag mAb (AE005)

    Immunofluorescence analysis of 293T-BCAT2-Flag-GFP-C cells using Mouse anti DDDDK-Tag mAb (AE005) at dilution of 1:100 (40x lens). Blue: DAPI for nuclear staining.
    ABclonal:Immunofluorescence - Mouse anti DDDDK-Tag mAb (AE005)}

    Immunofluorescence - Mouse anti DDDDK-Tag mAb (AE005)

    Immunofluorescence analysis of 293T-BCAT2-Flag-GFP-N cells using Mouse anti DDDDK-Tag mAb (AE005) at dilution of 1:100 (40x lens). Blue: DAPI for nuclear staining.
    ABclonal:Immunoprecipitation - Mouse anti DDDDK-Tag mAb (AE005)}

    Immunoprecipitation - Mouse anti DDDDK-Tag mAb (AE005)

    Immunoprecipitation of SERPINB1-Flag from 150 µg extracts of 293T cells transfected with a SERPINB1 expression vector containing a single N-terminal DDDDK-Tag was performed using 0.5 µg of Mouse anti DDDDK-Tag mAb (AE005). Mouse IgG isotype control (AC011) was used to precipitate the Control IgG sample. IP samples were eluted with 1X Laemmli Buffer. The Input lane represents 10 % of the total input. Western blot analysis of immunoprecipitates was conducted using Mouse anti DDDDK-Tag mAb (AE005) at a dilution of 1:5000.
    ABclonal:Flow CytoMetry - Mouse anti DDDDK-Tag mAb (AE005)}

    Flow CytoMetry - Mouse anti DDDDK-Tag mAb (AE005)

    Flow cytometry: 1X10^6 CHO cells (negative control, left) and CHO-Claudin18.2-Flag (Transfection, right) cells were intracellularly-stained with Mouse anti DDDDK-Tag mAb (AE005, 2 μg/mL, orange line) or Mouse isotype control (2 μg/mL, blue line), followed by PE Goat anti-Mouse pAb staining. Non-fluorescently stained cells were used as blank control (red line).
    ABclonal:Flow CytoMetry - Mouse anti DDDDK-Tag mAb (AE005)}

    Flow CytoMetry - Mouse anti DDDDK-Tag mAb (AE005)

    Flow cytometry: 1X10^6 CHO-Claudin18.2-Flag (Transfection, right) cells were intracellularly-stained with Mouse isotype control (2 μg/mL, left) or Mouse anti DDDDK-Tag mAb (AE005, 2 μg/mL, right), followed by PE Goat anti-Mouse pAb staining.

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

    Publishing research using AE005? Please let us know so that we can cite the reference in this datasheet.