PMY-2A4
Catalog Fields
Product Name/ID: PMY-2A4
Available to For-Profits: Yes
Alternate Antibody Name: clone 4A12
Gene Name:
Ab Isotype: MIgG2c
Gene Symbol:
Antibody Registry ID: AB_2619605
Clonality: Monoclonal
Immunogen: Puromycin (coupled to KLH) from Streptomyces alboniger bacterium. Chemical Composition: 3'-(alpha-Amino-p-methoxyhydrocinnamamido)-3'-deoxy-N,N-dimethyladenosine, 2HC1.
Clone:
Immunogen Sequence:
Myeloma Strain: SP2
Epitope Mapped: Yes
Antigen Name: puromycin
Epitope Location or Sequence:
Alternate Antigen Name:
Deposit Date: 3/9/2012
Antigen Molecular Weight: 0.5kDa
Depositor: Yewdell, Jonathan
Antigen Sequence:
Depositor Institution: National Institutes of Health
Antigen Species: Streptomyces alboniger
Depositor Notes: Fusion date: 07/21/2010. Fixation, 4% paraformaldehyde. The antigen puromycin inhibits translation by covalently associating with nascent chain within the ribosome. 2A4 allows puromycin detection in this condition. 2A4 also detects any puromycylated peptide. Medium: RPMI 1640 / Glutamine / Na pyruvate / 10% fetal bovine serum / Penicillin-streptomycin /gentamycin
Host Species: mouse
Hybridoma Cells Available (Non-Profit): Yes
Confirmed Species Reactivity: All
Additional Information: Puromycin is a structural analog of aminoacyl tRNAs that can be incorporated into nascent polypeptide chain and blocks elongation. This antibody is used to detect the subcellular localization of actively translating ribosomes by the ribopuromycylation method (RPM), and localizes puromycylated nascent chains on ribosomes immobilized by chain elongation inhibitors in living cells prior to processing for intracellular immunofluorescence and/or western blots. RRID:AB_2619605
Recommended Applications: ELISA, FACS, Immunofluorescence, Immunoprecipitation, Western Blot
All cell products contain the antimicrobial ProClin. Click here for additional information.
These hybridomas were created by your colleagues. Please acknowledge the hybridoma contributor and the Developmental Studies Hybridoma Bank (DSHB) in the Materials and Methods of your publications. Please email the citation to us.
For your Materials & Methods section:
PMY-2A4 was deposited to the DSHB by Yewdell, Jonathan (DSHB Hybridoma Product PMY-2A4)
Storage and Handling Recommendations
Although many cell products are maintained at 4°C for years without loss of activity, shelf-life at 4°C is highly variable. For immediate use, short term storage at 4°C up to two weeks is recommended. For long term storage, divide the solution into volumes of no less than 20 ul for freezing at -20°C or -80°C. The small volume aliquot should provide sufficient reagent for short term use. Freeze-thaw cycles should be avoided. For concentrate or bioreactor products, an equal volume of glycerol, a cryoprotectant, may be added prior to freezing.
Usage Recommendations
The optimal Ig concentration for an application varies by species and antibody affinity. For each product, the antibody titer must be optimized for every application by the end user laboratory. A good starting concentration for immunohistochemistry (IHC), immunofluorescence (IF), and immunocytochemistry (ICC) when using mouse Ig is 2-5 ug/ml. For western blots, the recommended concentration range of mouse Ig 0.2-0.5 ug/ml. In general, rabbit antibodies demonstrate greater affinity and are used at a magnitude lower Ig concentration for initial testing. The recommended concentrations for rabbit Ig are 0.2-0.5 ug/ml (IF, IHC and ICC) and 20-50 ng/ml (WB).
17 References
Initial Publication
Nuclear translation visualized by ribosome-bound nascent chain puromycylation.
Yewdell JW
The Journal of cell biology 197.1 (2012 Apr 2): 45-57.