Anti-MNK2 antibody (ab84345) at 1 µg/ml (in 5% skim milk / PBS buffer) + 721_B cell lysate at 10 µgSecondaryHRP conjugated anti-Rabbit IgG at 1/50000 dilution
ICC/IF image of ab84345 stained HeLa cells. The cells were 100% methanol fixed (5 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 (ab84345, 1µg/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.
ab84345 at 5 µg/ml staining MNK2 in Human Placenta tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
ab84345 at 5 µg/ml staining MNK2 in Human spleen tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
IHC image of ab84345 staining in Human breast cancer 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 ab84345, 5µg/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.
All lanes : Anti-MNK2 antibody (ab84345) at 1 µg/mlLane 1 : MarkerLane 2 : Zebrafish brain homogenate at 20 µgLane 3 : Zebrafish heart homogenate at 10 µgLane 4 : Zebrafish liver homogenate at 10 µgLane 5 : Zebrafish skeletal muscle homogenate at 10 µgLane 6 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µgSecondaryGoat polyclonal to Rabbit IgG – H&L – Pre-Adsorbed (HRP) at 1/6000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.