![Fatty Acid Binding Protein 5 was immunoprecipitated using 0.5mg A431 whole cell extract, 5µg of Rabbit polyclonal to 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, A431 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 ab84028.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 16kDa; Fatty Acid Binding Protein 5](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_2/17746_ab84028-215253-IPVI006ab8402820mMod.jpg)
Fatty Acid Binding Protein 5 was immunoprecipitated using 0.5mg A431 whole cell extract, 5µg of Rabbit polyclonal to 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, A431 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 ab84028.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 16kDa; Fatty Acid Binding Protein 5
All lanes : Anti-Fatty Acid Binding Protein 5 antibody (ab84028) at 1 µg/mlLane 1 : Skin (Human) Tissue Lysate - adult normal tissue (ab30166)Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 4 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell LysateLysates/proteins at 10 µg per lane.SecondaryGoat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
ICC/IF image of ab84028 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab84028, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) Hek293, HepG2, MCF-7 cells at 5µg/ml, and in 100% Methanol fixed (5 min) HeLa, Hek293, HepG2, MCF-7 cells at 5µg/ml.