Detection of FLAG proteins antibody

• Catalog Number: orb345396
• Category: Antibodies
• Description: Detection of FLAG proteins antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, IHC, WB
• Isotype: IgG
• Immunogen: This antibody was purified from whole rabbit serum prepared by repeated immunizations with the Enterokinase Cleavage Site (ECS) peptide DYKDDDDK (Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys) conjugated to KLH using maleimide. This antibody reacts with FLAG conjugated proteins.
• Concentration: 1.13mg/mL
• Dilution range: ELISA: 1:20,000 - 1:40,000, IHC: User Optimized, WB: 1:2,000 - 1:10,000
• Form/Appearance: Liquid (sterile filtered)
• Purity: This affinity purified antibody is directed against the FLAG motif and is useful in determining its presence in various assays. This polyclonal 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: rabbit antibody for the detection of FLAG conjugat
• Note: For research use only
• Application notes: This antibody 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. This antibody has been tested by ELISA and western blotting against both the immunizing peptide and FLAGä containing recombinant proteins. Although not tested, this antibody is likely functional for immunoprecipitation, immunocytochemistry, and other immunodetection techniques. The epitope tag peptide sequence was first derived from the 11-amino-acid leader peptide of the gene-10 product from bacteriophage T7. Now the most commonly used hydrophilic octapeptide is DYKDDDDK. Rockland Immunochemical's polyclonal antibody to detect FLAG conjugated proteins binds FLAG containing fusion proteins with greater affinity than the widely used monoclonal M1, M2 and M5 clones, and shows greater sensitivity in most assays. Affinity purification of the polyclonal antibody results in very low background levels in assays and low cross-reactivity with other cellular proteins.
• Expiration Date: 12 months from date of receipt.

References

Smith, Thomas H. et al. Protease-activated receptor-4 and purinergic receptor P2Y12 dimerize, co-internalize, and activate Akt signaling via endosomal recruitment of β-arrestin J Biol Chem, 292, 13867-13878 (2017)

PubMed: 28652403

Western blot analysis of shown to detect as little as 3 ng of amino-terminal FLAG tagged recombinant protein using Detection of FLAG proteins antibody

Western blot analysis of detect FLAG conjugated proteins detects both C terminal linked and N terminal linked FLAG tagged recombinant proteins using Detection of FLAG proteins antibody

Western Blot analysis of Lane 1: HEK293 lysate. Lane 2: HeLa Lysate. Lane 3: CHO/K1 Lysate. Lane 5: A431 Lysate Lane 6: Jurkat Lysate. Lane 7: NIH/3T3 Lysate. Lane 8: E-coli HCP Control. Lane 9: FLAG Positive Control Lysate. Lane 10: Red Fluorescent Prote

Western Blot analysis of of Lane 1: Opal Prestained Molecular Weight Marker. Lane 2: FLAG Positive Control Lysate(10ug) [+]. Lane 3: 12 Epitope GST Tagged Lysate (10ug) [+]. Lane 4: rGST protein (0.1µg) [-]. Lane 5: MBP protein (0.1ug) using Detection of