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Flag-Tag Rabbit Monoclonal Antibody

Cat Number: AB092
Conjugate: Unconjugated
Size: 100 ug
Clone: ARC5111-01
Concentration: 1mg/ml
Host: Rabbit
Isotype: IgG
Immunogen: Synthetic peptide. This information is considered to be commercially sensitive.
Reactivity: Species independent
Applications: WB 1:5000 - 1:20000 IP 0.5μg-4μg antibody for 200μg-400μg extracts of whole cells IF/ICC 1:300 - 1:2000 FC 1:50 - 1:200 ChIP 5 μg antibody for 10 μg-15 μg of Chromatin ChIP-seq 1:50 - 1:200 ELISA Recommended starting concentration is 1 μg/mL. Please optimize the concentration based on your specific assay requirements.
Molecular: 56kDa/50kDa/46kDa/68kDa
Purification: Affinity purification
Synonyms: DDDDK; DDDDK tag; DDDDK-tag; DDDDK-Tag
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.
Form: liquid
Buffer: PBS with 0.09% Sodium azide,0.05% BSA,50% glycerol,pH7.3.
Storage: Store at -20℃. Avoid freeze / thaw cycles.
Western blot analysis of lysates from wild type (WT) and 293T cells transfected with GSK3B Protein,BCAT2-N Protein,BCAT2-C Protein, using DDDDK-Tag antibody at 1:10000 dilution. Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) ) at 1:10000 dilution. Lysates/proteins: 25μg per lane. Blocking buffer: 3% nonfat dry milk in TBST. Detection: ECL West Pico Plus. Exposure time: 10s.

Western blot analysis of lysates from wild type (WT) and 293T cells transfected with GSK3B Protein,BCAT2-N Protein,BCAT2-C Protein, using DDDDK-Tag antibody at 1:10000 dilution.
Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) ) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL West Pico Plus.
Exposure time: 10s.

Western blot analysis of lysates from wild type (WT),293F transfected with COPB2-N Protein,COPB2-C Protein,using DDDDK-Tag antibody at 1:10000 dilution. Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L)at 1:10000 dilution. Lysates/proteins: 25μg per lane. Blocking buffer: 3% nonfat dry milk in TBST. Detection: ECL West Pico Plus). Exposure time: 30s.

Western blot analysis of lysates from wild type (WT),293F transfected with COPB2-N Protein,COPB2-C Protein,using DDDDK-Tag antibody at 1:10000 dilution.
Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L)at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL West Pico Plus).
Exposure time: 30s.

Chromatin immunoprecipitation was performed with 21.8 μg of cross-linked chromatin from 293T cells transfected with a GATA3 expression vector containing a single C-terminal DDDDK-Tag using 5 μg of Rabbit anti DDDDK-Tag. DNA libraries were prepared using Scale ssDNA-seq Lib Prep Kit for Illumina V2. The ChIP sequencing results indicate the enrichment pattern of DDDDK-Tag in the representative genomic region surrounding SH3RF3 gene.

Chromatin immunoprecipitation was performed with 21.8 μg of cross-linked chromatin from 293T cells transfected with a GATA3 expression vector containing a single C-terminal DDDDK-Tag using 5 μg of Rabbit anti DDDDK-Tag. DNA libraries were prepared using Scale ssDNA-seq Lib Prep Kit for Illumina V2. The ChIP sequencing results indicate the enrichment pattern of DDDDK-Tag in the representative genomic region surrounding SH3RF3 gene.