Rabbit anti DDDDK-Tag Polyconal Antibody (CABE004)
The Rabbit anti DDDDK-Tag Polyconal Antibody (CABE004) is a high-quality antibody developed for reliable detection and analysis of target proteins. 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.
This antibody is validated for use in WB, IF/ICC, IP applications and has demonstrated reactivity against Species independent samples.
Product Name:
Rabbit anti DDDDK-Tag Polyconal Antibody
SKU:
CABE004
Size:
100μL, 50μL
Reactivity:
Species independent
Conjugate:
Unconjugated
Immunogen:
Synthetic peptide. This information is considered to be commercially sensitive.
Tested Applications:
WBIF/ICCIP
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
Synonyms:
DDDDK, DDDDK tag, DDDDK-tag
Positive Sample:
293T
Observed MW:
48kDa
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.
Purification Method
Affinity purification
RRID
AB_2771921
Buffer Information
Store at -20℃. Avoid freeze / thaw cycles. Buffer: PBS containing 50% glycerol, preserved with proclin300 or sodium azide, pH 7.3.
Western blot analysis of lysates from wild type (WT) and 293T cells transfected with DDDDK-Tag using Rabbit anti DDDDK-Tag pAb (CABE004) 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 (AbGn00020). Exposure time: 1s.
Immunofluorescence analysis of 293T cells transfected with Flag-c(mCherry) and 293T cells transfected with Flag-N(mCherry) use Rabbit anti DDDDK-Tag pAb (CABE004) at dilution of 1:50 (40x lens). Secondary antibody: Cy3-conjugated Goat anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.
Immunoprecipitation analysis of 300 μg extracts of 293T-SERPINB1-Flag-GFP cells using 3 μg DDDDK-Tag pAb (CABE004). Western blot was performed from the immunoprecipitate using DDDDK-Tag pAb (CABE004) at a dilition of 1:50000.
