DYKDDDDK antibody

• Catalog Number: orb344453
• Category: Antibodies
• Description: DYKDDDDK antibody
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 29E4.G7
• Tested applications: ELISA, FC, IHC, WB
• Isotype: IgG2a
• Immunogen: This antibody was produced in mice by repeated immunizations with a synthetic peptide corresponding to the FLAG epitope tag peptide DYKDDDDK (Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys) conjugated to KLH using maleimide.
• Concentration: 1.0 mg/ml
• Dilution range: ELISA: 1:150,000 - 1:250,000, FC: User Optimized, IHC: 1:1,000-1:5,000, WB: 1:2,000-1:10,000
• Form/Appearance: Liquid (sterile filtered)
• Purity: This product is an IgG fraction antibody purified from ascites by Protein A chromatography followed by extensive dialysis against the buffer stated above. The purified antibody is directed against the FLAG motif and is useful in determining its presence in various assays where the epitope tag is present at either the amino or carboxy terminus of recombinant proteins. This monoclonal anti-FLAG tag antibody detects over-expressed proteins containing the FLAG epitope tag. In western blotting of bacterial extracts, the antibody does not cross-react with endogenous proteins.
• Conjugation: Unconjugated
• Storage: Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.
• Buffer/Preservatives: 0.01% (w/v) Sodium Azide
• Alternative names: mouse anti-FLAG tag, Enterokinase Cleavage Site (E
• Note: For research use only
• Application notes: Anti-FLAG antibody has been tested by ELISA and western blot and is optimally suited for monitoring the expression of FLAG tagged fusion proteins. As such, this antibody can be used to identify fusion proteins containing the FLAG epitope. The antibody recognizes the epitope tag fused to either the amino- or carboxy- termini of targeted proteins. The epitope tag peptide sequence was first derived from the 11-amino-acid leader peptide of the gene-10 product from bacteriophage T7. DYKDDDDK is the most commonly used hydrophilic octapeptide tag.
• Expiration Date: 12 months from date of receipt.

Monoclonal Antibody to detect FLAG™ conjugated proteins detects both C terminal linked and N terminal linked FLAG™ tagged recombinant proteins by western blot.

Twenty-four (24) clones were randomly selected and grown up from glycerol stocks by inoculating 0.5 mL 2xYT medium. Expression of recombinant proteins was induced by the addition of IPTG. Proteins were purified by nickel affinity chromatography and eluted in 40 µL. Samples were diluted 10-fold, transferred to nitrocellulose membrane and blotted using Mab-anti-FLAG™ antibody.