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Reassurance, Refunds & Replacements
If your product does not perform as described on this datasheet, we will refund or replace your product...
Read our guarantee »Publishing research using ab36722? Please let us know so that we can cite the reference in this datasheet
ab36722 has not yet been referenced specifically in any publications.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Immunohistochemistry (Paraffin-embedded sections) - MKK6 (phospho S207) antibody (ab36722)
Immunohistochemical analysis of paraffin embedded breast carcinoma, using ab36772 diluted 1:50. Left: Untreated Ab; Right: Ab pre-treated with synthesized phosphopeptide.
Western blot - MKK6 (phospho S207) antibody (ab36722)
All lanes : Anti-MEK6 (phospho S207) antibody (ab36722) at 1/500 dilution
Lane 1 : Extracts from untreated MDA-MB-435 cells
Lane 2 : Extracts from UV treated MDA-MB-435 cells
Lysates/proteins at 30 µg per lane.
Predicted band size : 37 kDa
Observed band size : 40 kDa (why is the actual band size different from the predicted?)
UV treatment protocol:
a. For adherent cells, cells were plated on 100-mm culture dishes with growth medium (basic medium containing 10% serum).
b. When the cells were at the log phase and 80-90% confluent, the culture media was drained. Cells were washed with fresh basic media (no serum) once.
c. Add 3ml fresh basic media per dish before irradiation.
d. Culture dishes were uncovered in a UV cross-linker. UV irradiation was carried out with 100 J/m2 (or with various other dosages).
e. Cells can be directly used to prepare cell lysis. If needed to reculture in some experiments, 3ml growth medium was added to per dish, and the cells were incubated at 37°C for 1~2 hours in a CO2 incubator.
For suspension cells, cells were collected by centrifugation (1000g, 10min) from cell supernatant from flasks. Resuspend the cells in 3ml of fresh basic media, and plate on a 100-mm plate for irradiation.
Immunocytochemistry/ Immunofluorescence-MEK6 (phospho S207) antibody(ab36722)
ICC/IF image of ab36722 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 (ab36722, 1µg/ml) overnight at +4ºC. 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.
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