All lanes : Anti-NTF97/Importin beta antibody (ab45938) at 2 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate Lane 3 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate (ab7179)Lane 4 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate Lysates/proteins at 15 µg per lane.SecondaryGoat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilutionPerformed under reducing conditions.
![NTF97/Importin beta was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to NTF97/Importin beta 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 ab45938.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 97kDa; NTF97/Importin beta](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_4/2420_NTF97Importin-beta-Primary-antibodies-ab45938-6.jpg)
NTF97/Importin beta was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to NTF97/Importin beta 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 ab45938.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 97kDa; NTF97/Importin beta
ab45938 (1/200) detecting NTF97 in HeLa cells (green). Cells were fixed with paraformaldehyde, permeabilised with 0.5% Triton X100/PBS and counterstained with DAPI in order to highlight the nucleus (red). Please refer to abreview for further experimental details.See Abreview
IHC image of ab45938 staining in human skin formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab45938, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.