Anti-ORP8 antibody [mAbcam 60110]

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-ORP8 antibody [mAbcam 60110] (AB60110)

Overlay histogram showing HEK293 cells stained with ab60110 (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 (ab60110, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HEK293 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

• WB

Project4406****

Western blot - Anti-ORP8 antibody [mAbcam 60110] (AB60110)

1% BSA was used as a blocking agent.

All lanes:

Western blot - Anti-ORP8 antibody [mAbcam 60110] (ab60110) at 5 µg/mL

Lane 1:

HEK293 (Human embryonic kidney cell line) Whole Cell Lysate at 10 µg

Lane 2:

THP1 (Human acute monocytic leukemia cell line) Whole Cell Lysate at 10 µg

Lane 3:

Human kidney tissue lysate - total protein (<a href='/en-us/products/unavailable/human-kidney-tissue-lysate-total-protein-ab30203'>ab30203</a>) at 10 µg

Secondary

All lanes:

Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 101 kDa

Observed band size: 101 kDa,44 kDa,55 kDa,80 kDa,85 kDa

true

Exposure time: 30s

• WB

Lab

Western blot - Anti-ORP8 antibody [mAbcam 60110] (AB60110)

We recommend using 3% milk as the blocking agent in Western Blot.

All lanes:

Western blot - Anti-ORP8 antibody [mAbcam 60110] (ab60110) at 5 µg/mL

Lane 1:

HEK293 (Human embryonic kidney cell line) Whole Cell Lysate at 10 µg

Lane 2:

THP1 (Human acute monocytic leukemia cell line) Whole Cell Lysate at 10 µg

Lane 3:

Human kidney tissue lysate - total protein (<a href='/en-us/products/unavailable/human-kidney-tissue-lysate-total-protein-ab30203'>ab30203</a>) at 10 µg

Secondary

All lanes:

Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/10000 dilution

Predicted band size: 101 kDa

Observed band size: 101 kDa

true

Exposure time: 8min