Our Abpromise guarantee covers the use of ab4137 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/250. Predicted molecular weight: 71 kDa. (using horseradish peroxidase or alkaline phosphatase).|
|ELISA||Use at an assay dependent concentration.|
|IHC-Fr||Use at an assay dependent concentration.|
|ICC||1/250. Frozen. (using horseradish peroxidase or alkaline phosphatase).|
|IP||1/300. PubMed: 19028678|
Image from Barros SA et al, J Biol Chem. 2009 Jan 30;284(5):2672-9. Epub 2008 Nov 21, Fig 2.Immunoprecipitation.Rat renal cortex from adult male Sprague-Dawley CD rats was isolated and suspended in CelLyticTM MT Mammalian Tissue Lysis/Extraction reagent containing complete protease inhibitor mixture. Tissues were then homogenized and centrifuged at 2,500 x g for 10 minutes at 4 °C. The supernatant was used for immunoprecipitation. To eliminate nonspecific binding, 1 ml of rat kidney lysate (3 mg of protein) was pre-cleared by incubation with 50µl of TrueBlotTM anti-rabbit Ig IP beads overnight at 4 °C. Cleared cell lysates were incubated with either 7µg of ab4137 or 1µg of polyclonal rabbit anti-rOAT3 for 2 hours at 4 °C. TrueBlot anti-rabbit Ig IP beads (30µl) was added and incubation was continued for 2 hours at 4 °C. Bound proteins were washed 7 times with increasing washing stringency IP buffers and eluted from beads by heating in 4 x NuPAGE LDS sample buffer for 10 minutes. The Invitrogen NuPAGE Novex Tris