DYKDDDDK Tag (FLAG) Antibody

SKU: orb344414

Description

Detection of FLAG antibody

Images & Validation

• Tested Applications: ELISA, WB
• Dilution Range: ELISA: 1:150,000 - 1:250,000, WB: 1:2,000 - 1:20,000
• Application Notes:
  Anti-FLAG has been tested by ELISA, SDS-Page, and western blot. Epitope tags are short peptide sequences that are easily recognized by tag-specific antibodies. Due to their small size, epitope tags do not affect the tagged protein’s biochemical properties. Most often sequences encoding the epitope tag are included with target DNA at the time of cloning to produce fusion proteins containing the epitope tag sequence. This allows anti-epitope tag antibodies to serve as universal detection reagents for any tag containing protein produced by recombinant means. This means that anti-epitope tag antibodies are a useful alternative to generating specific antibodies to identify, immunoprecipitate or immunoaffinity purify a recombinant protein. The anti-epitope tag antibody is usually functional in a variety of antibody-dependent experimental procedures. Expression vectors producing epitope tag fusion proteins are available for a variety of host expression systems including bacteria, yeast, insect and mammalian cells. Biorbyt produces anti-epitope tag antibodies against many common epitope tags including Myc, GST, GFP, 6X His, MBP, FLAG and HA. Biorbyt also produces antibodies to other tags including FITC, Rhodamine (TRITC), DNP and biotin.

Key Properties

• Antibody Type: Primary Antibody
• Host: Mouse
• Clonality: Monoclonal
• Isotype: IgG2a
• Clone No.: 29E4.G7
• 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.
• 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 & Handling

• 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.
• Form/Appearance: Liquid (sterile filtered)
• Buffer/Preservatives: Preservative: 0.01% (w/v) Sodium Azide. Stabilizer: None; Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
• Concentration: 1.026 mg/mL
• Expiration Date: 12 months from date of receipt.
• Dry Ice Shipping: Please note: This product requires shipment on dry ice. A dry ice surcharge will apply.
• Disclaimer: For research use only

Alternative Names

mouse anti-FLAG tag, Enterokinase Cleavage Site (ECS), mouse anti-DYKDDDDK, Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys

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.