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Synthetic peptide conjugated to KLH derived from within residues 50 - 150 of Mouse p19ARF.
(Peptide available as ab2659.)
Our Abpromise guarantee covers the use of ab80 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC||Use a concentration of 1 µg/ml. Fix with a 1:1 mixture of methanol:acetone at -20C for 10 mins (see image by Dr. David Bertwistle).|
|ELISA||Use at an assay dependent concentration.|
|IHC-P||Use a concentration of 1 µg/ml. PubMed: 11726500|
|WB||1/1000. Detects a band of approximately 19 kDa (predicted molecular weight: 17 kDa).Can be blocked with Mouse CDKN2A/p19ARF peptide (ab2659). Multiple bands with this antibody have frequently been reported. We believe that this is due to the different modification states of p19ARF. Nuclear extraction may be necessary in some samples.|
ab80 antibody was tested on NIH 3T3 cells (which have a deletion of the p19ARF gene) as a negative control (-) and NIH 3T3 cells engineered to express HA-tagged p19ARF as a positive control (+).
Cells grown on coverslips were fixed with a 1:1 mixture of methanol:acetone at -20C for 10 mins. After drying, rehydrating and blocking, the coverslips were incubated with ab80 at a concentration of 1ug/ml, diluted in PBS for 1 hour at room temperature. The antibody was then visualised using standard anti-rabbit secondary antibodies coupled to Texas Red.
ab80 at 1/300 staining mouse brain cells by ICC/IF.
Paraformaldehyde fixation, Triton X-100 permeabilization, followed by blocking with 3% BSA + 2% NGS for 1 hour at 20°C.
Incubation with the primary antibody was carried out for 1 hour at 20°C. The secondary antibody was an Alexa Fluor® conjugated Donkey anti-Rabbit polyclonal, diluted 1/400.
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