The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use 1µg for 106 cells. ab170191-Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
Use a concentration of 1 - 5 µg/ml. Predicted molecular weight: 51 kDa.
Use at an assay dependent concentration. The detection limit for recombinant tagged MNK1 is approximately 10 ng/ml when used as a capture antibody.
Use a concentration of 3 µg/ml. Antigen retrieval is not essential but may optimise staining.
Use a concentration of 10 µg/ml.
FunctionMay play a role in the response to environmental stress and cytokines. Appears to regulate transcription by phosphorylating EIF4E, thus increasing the affinity of this protein for the 7-methylguanosine-containing mRNA cap.
Sequence similaritiesBelongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. Contains 1 protein kinase domain.
Post-translational modificationsDual phosphorylation of Thr-250 and Thr-255 activates the kinase. Phosphorylation of Thr-385 activates the kinase.
Cellular localizationCytoplasm and Cytoplasm. Nucleus.
ab89223, at 10 µg/ml, staining MNK1 in paraformaldehyde fixed, permeabilised HeLa cell by Immunofluorescence.
Flow Cytometry-Anti-MNK1 antibody(ab89223)
Overlay histogram showing HeLa cells stained with ab89223 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab89223, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line). Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
References for Anti-MNK1 antibody (ab89223)
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