![GAPDH was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to GAPDH 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, Hela 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 ab97626.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 36kDa; GAPDH](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_2/22730_ab97626-195055-IPV020ab976261m.jpg)
GAPDH was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to GAPDH 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, Hela 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 ab97626.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 36kDa; GAPDH
All lanes : Anti-GAPDH antibody (ab97626) at 1/5000 dilutionLane 1 : 293T whole cell lysateLane 2 : A431 whole cell lysateLane 3 : H1299 whole cell lysateLane 4 : Hela whole cell lysateLysates/proteins at 30 µg per lane.
ab97626, at a 1/200 dilution, staining GAPDH in paraformaldehyde fixed A549. Lower image was merged with DNA probe.