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Western blot analysis of lysates from wild type (WT) and 293F cells transfected with Mouse anti DDDDK-Tag using Mouse anti DDDDK-Tag mAb at 1:20000 dilution incubated overnight at 4℃. | Secondary antibody: HRP-conjugated Goat anti-Mouse IgG (H+L) at 1:10000 dilution. | Lysates/proteins: 20 μg per lane. | Blocking buffer: 3% nonfat dry milk in TBST. | Detection: ECL Basic Kit | .Exposure time: 45s.
Western blot analysis of lysates from wild type (WT) and 293F cells transfected with Mouse anti DDDDK-Tag using Mouse anti DDDDK-Tag mAb at 1:20000 dilution incubated overnight at 4℃. | Secondary antibody: HRP-conjugated Goat anti-Mouse IgG (H+L) at 1:10000 dilution. | Lysates/proteins: 20 μg per lane. | Blocking buffer: 3% nonfat dry milk in TBST. | Detection: ECL Basic Kit | .Exposure time: 45s.
Western blot analysis of lysates from wild type (WT) and 293F cells transfected with Mouse anti DDDDK-Tag using Mouse anti DDDDK-Tag mAb at 1:20000 dilution incubated overnight at 4℃. | Secondary antibody: HRP-conjugated Goat anti-Mouse IgG (H+L) at 1:10000 dilution. | Lysates/proteins: 20 μg per lane. | Blocking buffer: 3% nonfat dry milk in TBST. | Detection: ECL Basic Kit | .Exposure time: 45s.
Confocal imaging of 293F cells transfected with DDDDK-Tag using Mouse anti DDDDK-Tag mAb followed by a further incubation with Cy3 Goat Anti-Mouse IgG (H+L) (Red). DAPI was used for nuclear staining (Blue). Objective: 100x.
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. Mouse IgG isotype control 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 at a dilution of 1:5000.
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 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).
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, followed by PE Goat anti-Mouse pAb staining.
Western blot analysis of lysates from wild type (WT) and 293F cells transfected with Mouse anti DDDDK-Tag using Mouse anti DDDDK-Tag mAb at 1:20000 dilution incubated overnight at 4℃. | Secondary antibody: HRP-conjugated Goat anti-Mouse IgG (H+L) at 1:10000 dilution. | Lysates/proteins: 20 μg per lane. | Blocking buffer: 3% nonfat dry milk in TBST. | Detection: ECL Basic Kit | .Exposure time: 45s.

Mouse anti DDDDK-Tag Recombinant Antibody (84-509)

CATALOG NUMBER: 84-509

Clonality:
Monoclonal
Tested Applications:
ELISA, FC (intra), IF/ICC, IP, WB
Host Species:
Mouse
Species Reactivity:
Species independent
Conjugate:
Unconjugated
Specifications
Host Species:
Mouse
Species Reactivity:
Species independent
Immunogen:
Synthetic peptide. This information is considered to be commercially sensitive.
Conjugate:
Unconjugated
Tested Applications:
ELISA, FC (intra), IF/ICC, IP, WB
Application Note:
WB,1:10000 - 1:80000, IF/ICC,1:200 - 1:800, 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.
purification:
Affinity purification
Clonality:
Monoclonal
Isotype:
IgG1,Kappa
physical-state:
Liquid
Buffer:
Buffer: PBS with 0.09% Sodium azide,50% glycerol,pH7.3.
Concentration:
Batch dependent
Storage Conditions:
Store at -20℃. Avoid freeze / thaw cycles.
Ncbi Official Symbol:
DDDDK
Additional Names:
DDDDK;DDDDK tag;DDDDK-tag
User Note:
Optimal dilutions for each application to be determined by the researcher.
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.

FOR RESEARCH USE ONLY

For additional information, visit ProSci’s Terms & Conditions Page.

Disclaimer:
This product is for research use only.

CATALOG NUMBER:

84-509

List Size:
100 uL

List Price:

$245.00

Datasheet
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