Anti-GOLPH3/MIDAS antibody

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-GOLPH3/MIDAS antibody (AB98023)

ICC/IF image of ab98023 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 (ab98023, 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. This antibody also gave a positive result in 4% PFA fixed (10 min) Hek293, HepG2 and MCF7 cells at 5µg/ml.

• WB

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Western blot - Anti-GOLPH3/MIDAS antibody (AB98023)

The predicted molecular weight of GOLPH3/MIDAS is 34-kDa. We are unsure why we are seeing an additional band in HeLa and A549 whole cell lysates. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.

All lanes:

Western blot - Anti-GOLPH3/MIDAS antibody (ab98023) at 1 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 2:

Lung (Human) Tissue Lysate at 10 µg

Lane 3:

A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate at 10 µg

Lane 4:

Western blot - Mouse lung normal tissue lysate - total protein (<a href='/en-us/products/tissue-lysates/mouse-lung-normal-tissue-lysate-total-protein-ab29297'>ab29297</a>) at 10 µg

Lane 5:

Lung (Rat) Tissue Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 34 kDa

Observed band size: 32 kDa,34 kDa

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Exposure time: 30s