CAC Antibody Collection
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Puromycin-specific
Puromycin is an aminonucleoside antibiotic derived from the bacterial strain Streptomyces alboniger. Structurally resembling the 3' end of aminoacyl-tRNAs, it can bind to translating ribosomes and cause premature chain termination. Classically, protein biosynthesis has been monitored using pulse-chase and flooding dose methods which both employ radioactive amino acid labels.| Product name (click for order info) | Cat No (click for datasheet) |
Host | Species specificity |
| Anti Puromycin mAb (Clone 3RH11) | CAC-PEN-MA001 | MS | - |
| Product name | Anti Puromycin mAb (Clone 3RH11) |
| Cat No | CAC-PEN-MA001 |
| Description | Analysis using puromycin immunodetection is an advantageous alternative to radioactive amino acid labeling and allows for the direct evaluation/quantification of translation using standard immunochemical methods. Features 1) Easy to evaluate and quantify protein biosynthesis 2) An alternative to the traditional pulse-chase method that relies on radioactive amino acids 3) Applicable to Western blot and ELISA analyses References: 1) Kelleher AR, Gordon BS, Kimball SR, Jefferson LS. Changes in REDD1, REDD2, and atrogene mRNA expression are prevented in skeletal muscle fixed in a stretched position during hindlimb immobilization. Physiol Rep. 2014 Feb 25;2(2):e00246. doi: 10.1002/phy2.246. eCollection 2014 Feb 1. PubMed PMID: 24744910; PubMed Central PMCID: PMC3966240. 2) Kelleher AR, Kimball SR, Dennis MD, Schilder RJ, Jefferson LS. The mTORC1 signaling repressors REDD1/2 are rapidly induced and activation of p70S6K1 by leucine is defective in skeletal muscle of an immobilized rat hindlimb. Am J Physiol Endocrinol Metab. 2013 Jan 15;304(2):E229-36. doi: 10.1152/ajpendo.00409.2012. Epub 2012 Nov 27. PubMed PMID: 23193052; PubMed Central PMCID: PMC3543567. |
| Host | Mouse |
| Species specificity | - |
| Figure 1 |  |
| Immunoblot Human fibroblasts (TIG-1 cells) were plated in 12-well plates and cultured till 80% confluent. Puromycin (final conc. 0 uM and 5 uM) was added to the medium of the cells 80 min before harvest. Wells were washed with PBS, followed by addition of 1mL buffer (20 mM Tris-HCl (pH8.0) + protease inhibitor) and then ultrasonic fragmentation. Lysate was centrifuged (4°C, 12,000 g, 20 min) and supernatant was collected for downstream assays (Western Blotting, ELISA). Loaded 15 ug/lane. Primary Antibody (anti puromycin [CAC-PEN-MA001]) 1:1,000 dilution (1 ug/mL). Secondary Antibody (anti mouse Ig-HRP, DAKO, Cat.No. P0161)1:2,000 dilution |