Monoclonal ANTI-FLAG® antibody produced in rabbit
SIGMA/F2555 - clone SIG1-25, ascites fluid
Synonym: Anti-ddddk; Anti-dykddddk
Product Type: Chemical
Immunofluorescence Whole extract of 293T cells expressing FLAG® tagged fusion protein were fixed and permeabilized with 3% to 4% paraformaldehyde followed by 0.5% Triton™ X-100. Fixed cells were stained with 1:250 Monoclonal ANTI-FLAG® antibody produced in Rabbit, Clone: SIG1-25 (Cat. No. F2555).
Immunofluorescence Whole extract of 293T cells expressing FLAG® tagged fusion protein were fixed and permeabilized with 3% to 4% paraformaldehyde followed by 0.5% Triton™ X-100. Fixed cells were stained with 1:250 Monoclonal ANTI-FLAG® antibody produced in Rabbit, Clone: SIG1-25 (Cat. No. F2555).
Immunoblotting Whole extract of human HEK-293T cells overexpressing N-terminal Flag tagged fusion protein was separated on SDS-PAGE and probed with Rabbit Monoclonal Anti-Flag Clone:SIG1-25 (Cat. No. F2555). The antibody was developed using Goat Anti-Rabbit IgG-Peroxidase (Cat. No. A0545) and a chemiluminescent substrate. Lanes 1. Antibody dilution 1:100 2. Antibody dilution 1:250
| antibody form | ascites fluid |
| antibody product type | primary antibodies |
| biological source | rabbit |
| clone | SIG1-25, monoclonal |
| conjugate | unconjugated |
| immunogen sequence | DYKDDDDK |
| isotype | IgG |
| Quality Level | 200  |
| shipped in | dry ice |
| storage temp. | −20°C |
| technique(s) | immunocytochemistry: 1:125-1:250 using transiently transfected cells expressing FLAG (sequence at the N-terminus)-tagged protein fixed with paraformaldehyde/Triton™ X-100 |
| indirect ELISA: suitable | |
| western blot: 1:250-1:500 using extracts of transiently transfected cells expressing FLAG (sequence at the N-terminus)-tagged protein |
| Application: | Monoclonal ANTI-FLAG® antibody produced in rabbit has been used in: • immunofluorescence • western blot analysis • immunocytochemistry • indirect ELISA Learn more product details in our FLAG® application portal. |
| Biochem/physiol Actions: | This monoclonal antibody recognizes the FLAG sequence at the N-terminus |
| General description: | Epitope tags provide a method to localize gene products in a variety of cell types, study the topology of proteins and protein complexes, identify associated proteins, and characterize newly identified, low abundance, or poorly immunogenic proteins when protein specific antibodies are not available. Tagging with the FLAG® peptide sequence may be done at the N-terminus, N-terminus preceded by a methionine residue, C-terminus, or at internal positions of the target protein. FLAG may also be placed in association with other tags. The small size of the FLAG tag or sequence and its high hydrophilicity tend to decrease the possibility of interference with the protein expression, proteolytic maturation, antigenicity, and function. The N-terminal FLAG® peptide sequence contains a unique enterokinase cleavage site allowing it to be completely removed from the purified fusion proteins. Cleavage catalyzed by Cu2+ ions of the C-terminal FLAG® peptide from a fusion protein has been reported. A sequence motif with five out of eight amino acid residues identical to the FLAG® peptide is found in both rat and mouse Mg2+ dependent protein b-phosphatase, as well as in the human and bovine enzyme. This rabbit ANTI-FLAG® antibody is produced from the hybridoma SIG1-25. The hybridoma is derived by the fusion of myeloma cells and splenocytes from rabbits immunized with the FLAG® peptide sequence conjugated to KLH. |
| Immunogen: | FLAG peptide sequence DYKDDDDK |
| Legal Information: | ANTI-FLAG is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Legal Information: | FLAG is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Legal Information: | Triton is a trademark of The Dow Chemical Company or an affiliated company of Dow |
| Physical form: | Provided as ascites fluid containing 15 mM sodium azide as a preservative. |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 3 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | −20°C |
| UNSPSC | 12352203 |