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Anti-FIVE-tag
Description
The FIVE-tag is derived from a peptide within the HI virus Nef protein. The sequence of the five amino acids comprising the FIVE-tag (PEYFK) is one of the most compact epitope tags available. Importantly, the monoclonal antibody directed against the FIVE-tag shows low background binding to other proteins even in high complexity samples such as whole cell lysates. The versatility of our monoclonal antibody in Western Blotting, immunoprecipitation, or immunofluorescence staining and the uncomplicated inclusion of the FIVE-tag coding sequence in any expression vector during PCR-based cloning make this antibody exceptionally useful for every laboratory.
Product
Immunogen:
Synthetic peptide (PEYFK) derived from the HI virus Nef protein
Antibody-type:
mouse monoclonal (clone TT1)
Isotype:
IgG1
Concentration:
1 mg/ml
Purification:
Protein G affinity chromatography
Supplied buffer:
PBS, pH 7.2, containing 50% glycerol.
Shipping and storage
Shipping:
Antibody is shipped in cold case
Storage:
Antibody is stable for 1 month at 4°C. For prolonged storage, the antibody should be stored at -20°C. Aliquot to avoid repeated freezing and thawing. At -20°C, the product is stable for at least 1 year from shipment.
Use
For research use only. Not for diagnostic or therapeutic purpose.
Applications
Western Blotting with the FIVE-tag antibody
Lysates from human HEK293 cells transiently
transfected with the empty vector (control; lane A)
or a eukaryotic expression vector encoding
a ~70 kDa protein either without (proteinX; lane B)
or with an amino-terminal FIVE-tag (FIVE-proteinX;
lane C) were separated by SDS-PAGE and
transfered onto a PVDF membrane.
The membrane was probed with the anti-FIVE-tag antibody (clone TT1; 1:1,000 dilution).
Bound antibody was visualized using horseradish
peroxidase-coupled Protein G (1:10,000 dilution)
and chemiluminescence detection.
Exposure time 30 seconds (upper panel).
Each lane contained equal amount of total protein.
Western Blotting: 1:500 – 1:2,000
Immunoprecipitation: 2 µg/sample
Immunofluorescence staining: 1:200
(paraformaldehyde-fixed cells and tissues)
Immunohistochemistry: 1:100
(cryosections)
Optimal dilutions are dependent on experimental conditions and should be determined by the user.
