R1481P

antibody from Acris Antibodies GmbH

Targeting: RAD9A RAD9

Western blot (Supportive data in Antibodypedia)

ELISA (Recommended by provider)

Antibody Data

Product number

R1481P

Provider

Acris Antibodies GmbH

Proper citation

Acris Antibodies GmbH Cat#R1481P, RRID:AB_1006849

Product name

anti RAD9A pSer1260

Antibody type

Polyclonal

Antigen

This affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to phosphorylated form of aa 1249-1263 of 1309 of yeast Rad9 protein conjugated to KLH.

Reactivity

Yeast

Host

Rabbit

Vial size

0.1 mg

Concentration

0.41 mg/ml (by UV absorbance at 280 nm)

Western blot

Supportive validation

Submitted by

Acris Antibodies GmbH (provider)

Main image

Experimental details

Figure 1. Immunoblotting. Affinity purified phosphospecific antibody to yeast Rad9 at pS1260 was used at a 1:200 dilution incubated overnight at 4°C to detect Rad9 by Western blot. Lanes were loaded with 50 ng each of recombinant GST fusion protein containing S. cerevisiae Rad9 (aa 991-1309 ~60 kDa) on a 4-20% Criterion gel for SDS-PAGE as follows: Lane 1-non-phosphorylated wild type yeast Rad9, Lane 2-in vitro phosphorylated wild type yeast Rad9, Lane 3-non-phosphorylated S1129A/S1260A double mutant Rad9, Lane 4 - in vitro phosphorylated S1129A/S1260A double mutant. Phosphorylation of Rad9 was by treatment with ATP and Rad53 kinase. Rad53 kinase autophosphorylates and appears cross reactive as it is detected on the blot as 90 and 110 kDa bands (asterisk). Detection occurred using a 1:5,000 dilution of IRDye TM800 conjugated Donkey anti-Rabbit IgG for 1h at room temperature. LICOR's Odyssey® Infrared Imaging System was used to scan and process the image. Other detection systems will yield similar results