Application
Immunoprecipitation
Sample
African Green Monkey Cell lysate - whole cell (CV1 cells)
Total protein in input
1e+007 cells
Specification
CV1 cells
Immuno-precipitation step
Protein A/G
Other product details
Dilution
1/150
Incubation time
2 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: lysis buffer (see details)
Western Blot antibody
Name
Non-Abcam antibody was used: anti-Axl
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/2000
Additional data
Additional Notes
Cells from a confluent 75cm flask were lysed in a lysis buffer containig: 1% Triton X-100, 50 mM Tris pH 7.5, 150 mM NaCl, 1 mM EDTA, 1 mM EGTA, 1 mM DTT, 1 mM PMSF, 0.1 mM Na-Vanadate, 20 mM NaF and protease inhibitor cocktail. lyaste were placed on ice for 20 min then centrifuged at 14,000 rpm for 15 min at 4C.
Lysate was split into 2 tubes, pre-cleared for 1 hr with 1% rabbit serum+protein A/G beads and IP was performed with 2 ug of anti-Axl (either ab89224 or ab54803) + A/G beads for 2 hrs on a rotary shaker at 4C. beads were washed twice in lysis buffer and extracted with 2Xsample buffer.
lane 1 - 10 ul input lysate
lane 2 - IP with 2 ug ab89224
lane 3 - IP with 2 ug ab54803
Lysate was split into 2 tubes, pre-cleared for 1 hr with 1% rabbit serum+protein A/G beads and IP was performed with 2 ug of anti-Axl (either ab89224 or ab54803) + A/G beads for 2 hrs on a rotary shaker at 4C. beads were washed twice in lysis buffer and extracted with 2Xsample buffer.
lane 1 - 10 ul input lysate
lane 2 - IP with 2 ug ab89224
lane 3 - IP with 2 ug ab54803
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.
Mr. Nir Drayman
Verified customer
提交于 May 11 2011