Items Tagged ‘Fero-Protein’

June 1st, 2016

p27 ELISA

By UNM CC UNM CC

S.R. — 1/2004 Procedure Dilute p27 monoclonal antibody 1:1000 (v/v) in Carbonate Coating Buffer. Add 100 µl/well and incubate o/n @ 4°C or 1 hr @37°C. Wash 3X with TBST (pour onto plate, empty into sink, hit onto towel 3x to clear wells). Block plate by adding 270 µl/well of BSA/TBS solution. Incubate @37°C for […]

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Tags: Fero-Protein


June 1st, 2016

Western Blot Protocol

By UNM CC UNM CC

M.Fero 12/2011 (Includes sample preparation, Bradford Assay, SDS-PAGE, semi-dry transfer, antibody staining and ECL development.) Materials See below. Harvesting Cells Prepare lysis buffer by adding fresh protease inhibitors (100x stocks listed below) to RIPA or TG buffer, plus 1/1000 vol. of PMSF (from a 200 mM stock).  Also add fresh phosphase inhibitors (from 100x stock) […]

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Tags: Fero-Protein


June 1st, 2016

Histone Kinase Assay

By UNM CC UNM CC

(M. Fero) 9/1/04 Protocol To 1.5 mL eppendorf tubes add: 200 µg of protein extract (see Western blot protocol for protein sample preps) q.s. to 300 µL with RIPA (with protease and phosphatase inhibitors). Primary antibody (amount determined emperically, approx 10x the concentration needed for westerns) Vortex and incubate on ice for 30 min. Meanwhile […]

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Tags: Fero-Protein