| 货号 | 4336-APC-050 |
| 别名 | PAR | 全称 | Poly [ADP-ribose] Polymer |
| 应用 | Western Blot(1:1000 dilution) |
| 目标/特异性 | This polyclonal antibody detects free PAR and poly-ribosylated proteins. |
| 使用方法 | Western Blot: 1:1000 dilution |
| 来源 | The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below. |
| 产品组分 |
| 供应商 | R&D Systems |
| 纯化方式 | Antigen Affinity-purified |
| 免疫原 | Poly(ADP-ribose) polymer |
| 标记 | Unconjugated |
| 背景 | Procedure for Immunoblotting using Peroxidase Detection: Blotting buffer: 12 mM Tris base, 96 mM Glycine, and 15% MeOH. Blocking solution: 5% (w/v) nonfat dry milk in TBS-0.1% Tween. Antibody solution: 5% (w/v) nonfat dry milk, in TBS-0.1% Tween. Transfer the electrophoresed proteins to a PVDF membrane and incubate the membranefor 1 hour at room temperature in blocking solution. Incubate the membrane overnight at 4°C in antibody solution containing a 1:1000 dilution ofanti-PAR rabbit polyclonal antibody. Empirical determination of primary antibodyconcentration will be required for optimal results. Wash the membrane at room temperature for 5 minutes with 3 changes of TBS-0.1%Tween. Changing the membrane containers often reduces background. Incubate the membrane at room temperature for 1 hour in antibody solution containing antirabbitconjugated to horseradish peroxidase. Empirical determination of secondary antibody concentration will be required for optimalresults. Wash the membrane for 5 minutes with 4 changes of TBS-0.1% Tween. Develop peroxidase reaction using chemiluminescence. |
| 运输条件 | Blue Ice |
| 存放说明 | -20℃ |
| 参考文献 |
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Detection of PAR/pADPr by Western Blot. Western blot analysis of Wehi cellsuntreated (H) and treated (T) with 25 μM Etoposidefor 4 hours at 37 °C. Cells were lysed in Tris-GlycineSDS sample buffer at the concentration of 1 x 107cells/ml, and 10 μl of clarified lysate were loadedper well of 4-20% Tris-Glycine gel. Proteins weretransferred onto an Immobilon FL membrane andribosylated proteins were detected using Trevigenspolyclonal anti-PAR antibody (cat# 4336-APC-050)followed by an IR680-conjugated secondary antibody(Licor). The membrane was scanned using anOdyssey Infrared Imaging System (Licor). |
