MLPH Antibody / Melanophilin (FY12882)

Immunofluorescent staining of Melanophilin/MLPH using anti-MLPH antibody (green). Melanophilin/MLPH was detected in an immunocytochemical section of cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 ug/ml rabbit anti-MLPH antibody overnight at 4oC. DyLight 488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37oC. The section was counterstained with DAPI nuclear stain (blue). Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of Melanophilin/MLPH using anti-MLPH antibody. Lane 1: human MCF-7 whole cell lysates, Lane 2: human whole cell lysates, Lane 3: human PC-3 whole cell lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-MLPH antibody at 0.25 ug/ml overnight at 4oC, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal was developed using enhanced chemiluminescent. MLPH western blot shows a predominant band at ~85 kDa with adjacent upper and lower species. Although the theoretical mass is ~66 kDa, melanophilin commonly migrates higher due to phosphorylation and its low-complexity scaffold architecture; neighboring bands likely reflect differential modification states and minor proteolysis.