Anti-PSMD7/Mov34 抗体 (ab11436)

All lanes : Anti-PSMD7/Mov34 antibody (ab11436) at 1 µg/ml

Lane 1 : K-652 whole cell lysate
Lane 2 : HepG2 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : Jurkat whole cell lysate
Lane 5 : HEK-293 whole cell lysate
Lane 6 : U-2 OS whole cell lysate
Lane 7 : A549 whole cell lysate

Lysates/proteins at 30 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate at 0.25 µg/ml

Western blot analysis was performed on whole cell extracts (30 µg lysate). The blots were probed with ab11436 at 1 ug/ml and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (0.25 µg/mL, 1:4000 dilution). A 40 kDa band corresponding to PSMD7 was observed across the cell lines tested. In addition, a ~50 kDa band was observed, which could possibly represent the 26s Proteasome Subunit S5B. Known quantity of protein samples were electrophoresed and resolved proteins were then transferred onto a nitrocellulose membrane. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate.

IHC image of ab11436 staining in human normal skin formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM 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 ab11436, 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.

ICC/IF image of ab11436 stained HepG2 cells. The cells were 4% formaldehyde 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 (ab11436, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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.

Western blot of PSDM7 in K562 cell lysate using ab11436.
Lane 1 is ab11436
Lane 2 is ab11436 pre-incubated with peptide