Application
Immunoprecipitation
Immuno-precipitation step
Other - The product itself has the antibody attached to the sepharose beads
Sample
Saccharomyces cerevisiae Cell lysate - whole cell (yeast cell lysate)
Specification
yeast cell lysate
Total protein in input
5 µg
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: IP lysis buffer (Tris-Cl pH7.4,
Additional data
Additional Notes
In the experiment the protein of interest is tagged with a GFP tag and expressed with a constitutive promoter. The 4 OD of cells were lysed with glass beads (lysis buffer containing MOPS, SDS, Urea and EDTA) and the lysate was separated. Subsequently the GFP antibody coupled sepharose beads were added (20ul slurry). This was incubated overnight. Next day the beads were washes twice with the lysis buffer and SDS sample buffer was added and heated at 65 C. Lane 1 is the Input, Lane 2 is the unbound fraction, Lane 3 is the IP. The western blotting with anti-GFP antibody showed a specific band at 120kD.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
Abcam user community
Verified customer
投稿 Mar 26 2014