ANTI-FLAG® antibody, Rat monoclonal
SIGMA/SAB4200071 - clone 6F7, purified from hybridoma cell culture
Synonym: Anti-ddddk; Anti-dykddddk
MDL Number: MFCD02262912
Product Type: Chemical
Immunoprecipitation Rat Anti-FLAG Clone: 6F7 (Cat. No. SAB4200071) was used to immunoprecipitate BACE1 from a whole extract of HEK-293T cells overexpressing human BACE1-FLAG-tagged protein. Detection antibody: Rabbit Anti-FLAG (Cat. No. F7425) followed by Monoclonal Anti-Rabbit IgG native-peroxidase (Cat. No. R3155) and a chemiluminescent substrate. Lanes 1. 10 μg antibody 2. 2.5 μg antibody 3. Negative control: 10 μg nonrelevant antibody
Immunoblotting Whole extract of human HEK-293T cells overexpressing human Rab25-FLAG was separated on SDS-PAGE and probed with Rat Anti-FLAG Clone: 6F7 (Cat. No. SAB4200071). The antibody was developed using Rabbit Anti-Rat IgG-Peroxidase (Cat. No. A9542) and a chemiluminescent substrate. Lanes 1. 1 μg/mL antibody 2. Negative control: without first antibody
Immunoblotting N-terminal-FLAG-tagged BAP (Cat. No.P7582) (lanes 1-4) and C-terminal-FLAG-BAP (Cat. No. P7457) (lanes 5-8) proteins were separated on SDS-PAGE and probed with 2 μg/mL Rat Anti-FLAG Clone: 6F7 (Cat. No. SAB4200071). The antibody was developed using Rabbit Anti-Rat IgG-Peroxidase (Cat. No. A9542) and a chemiluminescent substrate. Lanes 1, 4. 50 ng protein 2, 5. 25 ng protein 3, 6. 10 ng protein 4, 8. 5 ng protein
| antibody form | purified from hybridoma cell culture |
| purified immunoglobulin | |
| antibody product type | primary antibodies |
| biological source | rat |
| clone | 6F7, monoclonal |
| conjugate | unconjugated |
| form | buffered aqueous solution |
| immunogen sequence | (DYKDDDDK) |
| isotype | IgG1 |
| shipped in | dry ice |
| species reactivity | all |
| storage temp. | −20°C |
| technique(s) | immunoprecipitation (IP): 2.5-5.0 μg using lysates of transiently transfected cells expressing C-terminal-FLAG-tagged protein |
| western blot: 0.5-1.0 μg/mL using extracts of transiently transfected cells expressing C-terminal-FLAG-tagged protein |
| Application: | ANTI-FLAG® antibody, Rat monoclonal has been used in: • chromatin immunoprecipitation (ChIP) • western blotting • coimmunoprecipitation • flow cytometric analysis |
| Application: | Browse additional application references in our FLAG® Literature  portal. |
| Application: | Learn more product details in our FLAG® application portal. |
| General description: | Monoclonal Anti-FLAG® (rat IgG1 isotype) is derived from the hybridoma 6F7 produced by the fusion of mouse myeloma cells and splenocytes from rat immunized with the FLAG® peptide. The antibody is purified from culture supernatant of hybridoma cells grown in a bioreactor. Monoclonal Anti-FLAG® recognizes N-terminal, C-terminal and internal Flag-tagged fusion proteins. The product is especially recommended for identifying C-terminal FLAG®-tagged fusion proteins. 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 associationith 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 of the C-terminal FLAG® peptide from a fusion protein catalyzed by Cu2+ ions 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. |
| Immunogen: | FLAG peptide 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 |
| Physical form: | Solution in 0.01 M phosphate buffered saline, pH 7.4, containing 15 mM sodium azide. |
| RIDADR | NONH for all modes of transport |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | −20°C |
| UNSPSC | 12352203 |