DYKDDDDK (FLAG tag) antibody (Agarose)

• Catalog Number: orb344560
• Category: Antibodies
• Description: Mouse monoclonal antibody against DYKDDDDK (FLAG tag) conjugated to Agarose.
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 29E4.G7
• Tested applications: IP, WB
• Isotype: IgG2a
• Immunogen: Anti-DYKDDDDK Affinity Gel 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.
• Concentration: 0.99 mg
• Dilution range: IP: 10 µL resin binds >1 µg FLAG fusion protein
• Form/Appearance: Suspension of agarose beads
• Purity: Anti-DYKDDDDK Affinity Gel is a purified mouse IgG2a monoclonal antibody coupled to activated agarose. This product is intended for purification of proteins containing the FLAG epitope tag sequence. Binding Specificity: Anti-DYKDDDDK Affinity Gel binds the FLAG epitope tag sequence (Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys) fused to the amino terminal, carboxy terminal or internal locations of targeted recombinant proteins expressed in transfected or transformed cells. D-Y-K-D-D-D-D-K peptide (p/n 000-000-383) is recommended for competitive elution to recover fusion protein (see protocol).
• Conjugation: Agarose
• Storage: Store vial at 4°C prior to opening.
• Buffer/Preservatives: 0.01% (w/v) Sodium Azide
• Alternative names: mouse anti-FLAG tag agarose gel, mouse anti-DYKDDD
• Note: For research use only
• Application notes: Anti-DYKDDDDK Affinity Gel has been tested by SDS-Page, IP, and western blot and is optimally suited for immunoprecipitation and purification of FLAG tagged fusion proteins. Anti-DYKDDDDK Affinity Gel antibody recognizes the FLAG epitope tag fused to either the amino- or carboxy- terminal ends or an internal location of targeted fusion 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. Use D-Y-K-D-D-D-D-K peptide (p/n 000-000-383) for competitive elution to recover fusion protein (see protocol). Anti-FLAG is a registered trademark of Sigma-Aldrich. Refer to the Protocol for complete instructions for use including preferred buffers for elution. Do not use buffers that may denature the anti-DYKDDDDK antibody.
• Expiration Date: 12 months from date of receipt.

SDS-PAGE of Anti-DYKDDDDK (FLAG® tag) Affinity Gel. Lane 1: Cell lysate before purification. Lane 2: Flow through (used cell lysate). Lane 3: Purified DYKDDDDK (FLAG® tag) recombinant protein (arrowhead). Load: (6 µL per lane). Predicted/Observed size: 70 kDa for DYKDDDDK tagged recombinant protein.

SDS-PAGE of Anti-DYKDDDDK (FLAG® tag) Affinity Gel. Lane 1: crude lysate containing over-expressed DYKDDDK-tagged recombinant protein. Lane 2: unbound flow-through of consisting of endogenous E.coli proteins. Lane 3: enriched recombinant protein. Load: 5 µg protein, 15 µL flow through. Predicted/Observed size: 70 kDa for DYKDDDDK tagged recombinant protein.

SDS-PAGE of Anti-DYKDDDDK (FLAG® tag) Affinity Gel. Lane 1: DYKDDDDK tagged recombinant protein. Lane 2: flow through. Lane 3: wash. Lane 4: eluted fraction one. Lane 5: eluted fraction two. Lane 6: eluted fraction three. Lane 7: blank. Lane 8: molecular weight markers. Load: 5 µg protein lane 1, 15 µL per each other lane. Predicted/Observed size: 135 kDa for DYKDDDDK tagged recombinant protein.