![EEF1A2 was immunoprecipitated using 0.5mg MCF7 whole cell extract, 5µg of Rabbit polyclonal to EEF1A2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).The antibody was incubated under agitation with Protein G beads for 10min, MCF7 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab60283.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 50kDa; EEF1A2](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_2/11796_ab60283-171836-IPV019ab602834m.jpg)
EEF1A2 was immunoprecipitated using 0.5mg MCF7 whole cell extract, 5µg of Rabbit polyclonal to EEF1A2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).The antibody was incubated under agitation with Protein G beads for 10min, MCF7 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab60283.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 50kDa; EEF1A2
All lanes : Anti-EEF1A2 antibody (ab60283) at 1 µg/mlLane 1 : Brain (Human) Tissue Lysate - adult normal tissue (ab29466)Lane 2 : Heart (Human) Tissue Lysate - adult normal tissue (ab29431)Lane 3 : Skeletal Muscle (Human) Tissue Lysate - adult normal tissue (ab29330)Lane 4 : JEG-3 (Human placental choriocarcinoma cell line) Whole Cell Lysate Lane 5 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate Lane 6 : MDA-MB-361 (Human breast adenocarcinoma cell line) Whole Cell Lysate Whole Cell Lysate Lane 7 : MDA-MB-231 (Human breast adenocarcinoma cell line) Whole Cell Lysate Lysates/proteins at 10 µg per lane.SecondaryGoat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.