PAR Monoclonal Antibody Affinity Purified

形态 ：液体

克隆性 ：多克隆

宿主 ：兔

规格 ：50µl

特异识别2-50单位长度的PAR聚合酶，但不能识别RNA和DNA相关结构、ADP核糖酶单体、NAD+或者其他核酸单体，用于检测核糖基化蛋白。
适用于Elisa，WB，IHC-in situ和免疫纯化；可以作为Elisa和WB分析PARP-PAR的阳性对照。

More Details about

Description: An affinity purified rabbit polyclonal antibody raised against poly(ADP-ribose) (PAR) polymer. Trevigen’s anti-PAR polyclonal antibody can be used to detect ribosylated proteins by immunodetection. Trevigen’s PARP treated control protein (cat# 4500-10-P) and PAR polymer (cat# 4336-100-01) may be used as positive controls.
Physical State: This antibody is an affinity purified IgG fraction in 1X PBS, containing 50% glycerol.
Immunogen: Poly(ADP-ribose) polymer.
Specificity: This polyclonal antibody detects free PAR and poly-ribosylated proteins.
Storage: -20°C (manual defrost freezer).
Applications: For western and dot blotting, an antibody dilution of 1:1000 is recommended.
For ELISA a 1:4000 antibody dilution is recommended. Empirical determination of antibody dilutions will be required for optimum results.

Cell Lysates for Western Blotting:
To prepare total cell lysates, cells are solubilized in 1X SDS gel sample buffer (63 mM Tris-HCl, pH = 6.8; 10% glycerol; 2% SDS, 2.5% β-mercaptoethanol, and 0.0025% bromophenol blue) at 1 x 107 cells per ml. The extracts are heated in a boiling water bath for 5 minutes, vortexed for 1 minute at the maximum speed and centrifuged at 10,000 x g for 10 minutes at room temperature. Electrophorese on 4-20% Tris-Glycine SDS-PAGE gels.

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 membrane for 1 hour at room temperature in blocking solution.
Incubate the membrane overnight at 4°C in antibody solution containing a 1:1000 dilution of anti-PAR rabbit polyclonal antibody. Empirical determination of primary antibody concentration 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 antirabbit conjugated to horseradish peroxidase.
Empirical determination of secondary antibody concentration will be required for optimal results.
Wash the membrane for 5 minutes with 4 changes of TBS-0.1% Tween. Develop peroxidase reaction using chemiluminescence.