All lanes : Anti-Chromodomain helicase DNA binding protein 5 antibody (ab66516) at 1 µg/mlLane 1 : TE 671 (Human Rhabdomyosarcoma) Whole Cell LysateLane 2 : Hela Whole Cell Lysate - Hydroxyurea Treated (48hr, 2uM)Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell LysateLane 4 : Hela Whole Cell Lysate - Bleomycin Treated (20U/ml)Lane 5 : Liver (Human) Tissue Lysate - adult normal tissue (ab29889)Lane 6 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell LysateLane 7 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 8 : JEG-3 (Human placental choriocarcinoma cell line) Whole Cell LysateLysates/proteins at 10 µg per lane.SecondaryGoat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilutionPerformed under reducing conditions.
All lanes : Anti-Chromodomain helicase DNA binding protein 5 antibody (ab66516) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Nuclear Lysate (ab14844)Lysates/proteins at 10 µg per lane.SecondaryGoat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
![Chromodomain helicase DNA binding protein 5 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Chromodomain helicase DNA binding protein 5 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 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab66516.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 240kDa: Chromodomain helicase DNA binding protein 5; non specific - 58kDa: We are unsure as to the identity of this extra band.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_1/28665_Chromodomain-helicase-DNA-binding-protein-5-Primary-antibodies-ab66516-6.jpg)
Chromodomain helicase DNA binding protein 5 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Chromodomain helicase DNA binding protein 5 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 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab66516.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 240kDa: Chromodomain helicase DNA binding protein 5; non specific - 58kDa: We are unsure as to the identity of this extra band.