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EMD Millipore
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25 May 2012
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Recognizes the ~19 kDa p19ARF protein in whole cell lysates of mouse embryonic fibroblasts (MEF).
| Product information | |||
|---|---|---|---|
| Host | Rat | ||
| Isotype | IgG2b | ||
| Immunogen | a synthetic peptide corresponding to amino acids 54-75 of mouse p19ARF | ||
| Clone | 5-C3-1 | ||
| Form | Liquid | ||
| Formulation | In PBS. | ||
| Species reactivity | mouse | ||
| Preservative | ≤0.1% sodium azide | ||
| Applications |
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| Avoid freeze/thaw | Yes | ||
| Positive control | Mouse embryonic fibroblasts (MEF) | ||
| Store and ship information | |||
|---|---|---|---|
| Storage | -20°C | ||
| Ship |
Blue Ice Only
Irritant |
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| Data | |||
|---|---|---|---|
 Detection of mouse p19<sup>ARF</sup> by immunoblotting. Samples: Whole cell lysate (30 μg) from MEF (lanes 1,3) and NIH3T3 (lanes 2,4) cells. Primary antibody: Anti-p19<sup>ARF</sup> (54-75) Rat mAb (12-A1-1) (Cat. No. CB1013) (3 μg/ml) (lanes 1,2) or Anti-p19ARF (54-75) Rat mAb (5-C3-1) (Cat. No. CB1012) (3 μg/ml) (lanes 3,4). Detection: chemiluminescence.  A. Detection of mouse p19<sup>ARF</sup> by immunoblotting. Samples: Whole cell lysate (25 µg) from NIH3T3 cells that have deleted the Arf gene (lane 1) and wild type mouse embryo fibroblasts (MEFs) at passage 6, which express p19<sup>ARF</sup> (lane 2). Primary antibody: Anti-p19<sup>ARF</sup> (54-75) Rat mAb (5-C3-1) (Cat. No. CB1012) (3 µg/ml). Detection: chemiluminescence. B. Detection of mouse p19<sup>ARF</sup> by immunoprecipitation. Samples: Whole cell lysates (250 µg) from NIH3T3 cells that have deleted the Arf gene (lane 5) and NIH3T3 cells expressing p19<sup>ARF</sup> with a HA-tag (lanes 3,4). Antibodies for IP: Normal rat IgG (negative control; lane 3) and Anti-p19<sup>ARF</sup> (54-75) Rat mAb (5-C3-1) (Cat. No. CB1012). Conditions for immunoblotting: Samples: Whole cell lysate (25 µg, equivalent to 10% of above immunoprecipitations) from NIH3T3 cells that have deleted the Arf gene (lane 1), NIH3T3 cells expressing p19ARF with a HA-tag (lane 2), and immunoprecipitated proteins as indicated above. Primary antibody: Anti-HA. Detection: chemiluminescence. C. Detection of mouse p19<sup>ARF</sup> by immunocytochemistry. Samples: Wild type mouse embryo fibroblasts (MEF) at passage 5 (panels 1,2) and NIH3T3 cells, which have deleted the Arf gene (panels 3,4), fixed in methanol/acetone. Primary antibody: Anti-p19<sup>ARF</sup> (54-75) Rat mAb (5-C3-1) (Cat. No. CB1012). Detection: Anti-Rat IgG secondary antibody, fluorescent conjugate (panels 1,3) and counterstained with Hoechst dye (panels 2,4). D. Detection of mouse p19<sup>ARF</sup> by staining paraffin sections. Sample: Wild-type mouse testis. Primary antibody: Anti-p19<sup>ARF</sup> (54-75) Rat mAb (5-C3-1) (Cat. No. CB1012) (green). Detection: fluorescence with DAPI counterstain (blue). |
