Biolegend, 624451, Ultra-LEAF™ Purified anti-DYKDDDDK Epitope Tag Antibody, 25μg

The DYKDDDDK tag, commonly referred to as Sigma®'s FLAG® Tag, is often used as a protein modification in order to simplify the labeling and detection of proteins. This unique amino acid sequence allows for specific antibody detection in western blotting and immunocytochemistry. Due to the short sequence, this modification is not likely to affect the structure or function of the modified proteins.
25μg
Verified Reactivity: Epitope tag
Reported Reactivity: Species independent
Antibody Type: Monoclonal
Host Species: Rat
Immunogen: DYKDDDDK peptide conjugated to KLH
Formulation: 0.2 µm filtered in phosphate-buffered solution, pH 7.2, containing no preservative.
Endotoxin Level: Less than 0.01 EU/µg of the protein (< 0.001 ng/µg of the protein) as determined by the LAL test.
Preparation: The Ultra-LEAF™ (Low Endotoxin, Azide-Free) antibody was purified by affinity chromatography.
Concentration: The antibody is bottled at the concentration indicated on the vial, typically between 2 mg/mL and 3 mg/mL. To obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.
Storage & Handling: The antibody solution should be stored undiluted between 2°C and 8°C. This Ultra-LEAF™ solution contains no preservative; handle under aseptic conditions.
Application: WB - Quality tested ICC, IP - Verified
Recommended Usage: Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested use of this reagent is 0.01 - 0.25 µg/mL. For immunocytochemistry, a concentration range of 0.05 - 1.0 μg/mL is recommended. For immunoprecipitation, the suggested use of this reagent is 2.5 µg/test. It is recommended that the reagent be titrated for optimal performance for each application.
Application Notes: W22083A is not suitable for ICFC application. For ICC, fixation permeabilization with 100% ice-cold methanol, or with 4% PFA followed by 100% ice-cold methanol is recommended at the indicated concentration. Fixation/permeabilization with 4% PFA followed by 0.5% Triton-X results in dimmer staining, with an optimal concentration in the 0.1 - 0.5 µg/mL range. For use of this antibody (clone W22083A) in Western blot, this antibody has been demonstrated to have approximately 25% better sensitivity than commonly used anti-DYKDDDDK clone L5. For use of this antibody (clone W22083A) in ICC, this antibody has increased background staining compared to commonly used anti-DYKDDDDK (clone L5).
RRID: AB_3083222 (BioLegend Cat. No. 624451) AB_3083222 (BioLegend Cat. No. 624452)
Structure: Epitope tag
Biology Area: Protein Purification, Western Blot Controls
Antigen References: Rogers EJ, et al. 2019. Front Mol Neurosci. 12:43. Li Y, et al. 2008. J Biol Chem. 283:22443-22456.
Gene ID: NA
UniProt: View information about Epitope tag on UniProt.org
Clone: W22083A
Regulatory Status: RUO
Other Names: FLAG, FLAG tag
Isotype: Rat IgG2b, κ
Q: Do you guarantee that your antibodies are totally pathogen free?
A: BioLegend does not test for pathogens in-house aside from the GoInVivo™ product line. However, upon request, this can be tested on a custom basis with an outside, independent laboratory.
Q: Does BioLegend test each Ultra-LEAF™ antibody by functional assay?
A: No, BioLegend does not test Ultra-LEAF™ antibodies by functional assays unless otherwise indicated. Due to the possible complexities and variations of uses of biofunctional antibodies in different assays and because of the large product portfolio, BioLegend does not currently perform functional assays as a routine QC for the antibodies. However, we do provide references in which the antibodies were used for functional assays and we do perform QC to verify the specificity and quality of the antibody based on our strict specification criteria.
Q: Does BioLegend test each Ultra-LEAF™ antibody for potential pathogens?
A: No, BioLegend does not test for pathogens in-house unless otherwise indicated.  However, we can recommend an outside vendor to perform this testing as needed.
Q: Have you tested this Ultra-LEAF™ antibody for in vivo or in vitro applications?
A: We don't test our antibodies for in vivo or in vitro applications unless otherwise indicated. Depending on the product, the TDS may describe literature supporting usage of a particular product for bioassay. It may be best to further consult the literature to find clone specific information.