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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H, A | WB, FACS, IF, ICC | M | Purified | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | MABE342 |
| Description | Anti-Puromycin Antibody, clone 4G11 |
| Background Information | Puromycin is an aminonucleoside antibiotic, derived from the Streptomyces alboniger bacterium, that functions as a protein synthesis inhibitor that blocks translation through premature chain termination in the ribosome. Monoclonal antibodies to puromycin may be used with standard immunochemical methods to directly monitor translation, a method known as surface sensing of translation (SUnSET). Part of the molecule resembles the 3 end of the aminoacylated tRNA, making it useful for protein translation analysis. Puromycin induces DNA fragmentation in thymocytes and in human HL-60 leukemia cells. |
| Product Information | |
|---|---|
| Format | Purified |
| Control |
|
| Presentation | Purified mouse monoclonal IgG1κ in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Anti-Puromycin antibody, clone 4G11 detects puromycin incorporated into protein. Monoclonal antibodies to puromycin may also be used with standard immunochemical methods. |
| Key Applications |
|
| Application Notes | Immunocytochemistry Analysis: A 1:2,000 dilution from a representative lot detected Puromycin-incorporated neosynthesized proteins in HeLa cells treated with Puromycin. However, a decrease in signal is observed with the addition of Cycloheximide, an inhibitor of protein biosynthesis in eukaryotic organisms. Western Blotting Analysis: A representative lot from an independent laboratory detected Puromycin-incorporated neosynthesized proteins in WB (David, A., et al. (2012). J Cell Biol. 197(1):45-57.; Schmidt, E., K., et al. (2009). Nat Methods. 6(4):275-277.). Immunofluorescence Analysis: A representative lot from an independent laboratory detected Puromycin-incorporated neosynthesized proteins in IF (David, A., et al. (2012). J Cell Biol. 197(1):45-57.; Schmidt, E., K., et al. (2009). Nat Methods. 6(4):275-277.). Fluorescence Activated Cell Sorting Analysis: A representative lot from an independent laboratory detected Puromycin-incorporated neosynthesized proteins in FACS (David, A., et al. (2012). J Cell Biol. 197(1):45-57.; Schmidt, E., K., et al. (2009). Nat Methods. 6(4):275-277.). Alexa Fluor® is a registered trademark of Life Technologies®. |
| Biological Information | |
|---|---|
| Immunogen | Puromycin from Streptomyces alboniger |
| Clone | 4G11 |
| Host | Mouse |
| Isotype | IgG1κ |
| Species Reactivity |
|
| Species Reactivity Note | Demonstrated to react with Human test sample, preincubated with Puromycin. Predicted to react with all species when test sample is incubated with Puromycin. |
| Antibody Type | Monoclonal Antibody |
| Purification Method | Protein G Purified |
| Molecular Weight | Puromycin is incorporated in neosynthesized proteins. In the presense of Puromycin only, this antibody detects Puromycin-incorporated neosynthesized proteins at multiple molecular weights. However, a decrease in signal is observed in the co-presense of Cycloheximide, an inhibitor of protein biosynthesis in eukaryotic organisms. |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Evaluated by Western Blotting in HEK293 cell lysates treated with Puromycin and Cyclohexamide, or with Puromycin only. Western Blotting Analysis: A 1:12,500 dilution of this antibody detected Puromycin-incorporated neosynthesized proteins in HEK293 cell lysates treated with Puromycin only. This antibody also detected small mounts of puromycin in HEK293 cells treated with Puromycin and Cyclohexamide. |
| Usage Statement |
|
| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Stable for 1 year at 2-8°C from date of receipt. |
| Packaging Information | |
|---|---|
| Material Size | 100 µL |