Anti-PMP70 antibody

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PMP70 antibody (AB85550)

IHC image of ab85550 staining in Human Cervical carcinoma 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 ab85550, 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

Lab

Immunocytochemistry/ Immunofluorescence - Anti-PMP70 antibody (AB85550)

ab85550 staining PMP70 in HepG2 cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab85550 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

• WB

Unknown

Western blot - Anti-PMP70 antibody (AB85550)

The 73 kDa band observed is comparable to the molecular weight seen with other commercially available antibodies to Rat ATP-binding cassette sub-family D member 3 (PMP70).

All lanes:

Western blot - Anti-PMP70 antibody (ab85550) at 1 µg/mL

Lane 1:

Liver (Rat) Tissue Lysate at 10 µg

Lane 2:

Kidney (Rat) Tissue Lysate at 10 µg

Lane 3:

Human liver tissue lysate - total protein (<a href='/en-us/products/unavailable/human-liver-tissue-lysate-total-protein-ab29889'>ab29889</a>) at 10 µg

Lane 4:

HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 5:

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

Secondary

All lanes:

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

Predicted band size: 75 kDa

Observed band size: 25 kDa,28 kDa,37 kDa,50 kDa,70 kDa

true

Exposure time: 2min

• WB

Supplier Data

Western blot - Anti-PMP70 antibody (AB85550)

Lanes 1 - 4 : Merged signal (red and green). Green - ab85550 observed at 75 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab85550 was shown to specifically react with PMP70 in wild-type HAP1 cells as signal was lost in ABCD3 (PMP70) knockout cells. Wild-type and ABCD3 (PMP70) knockout samples were subjected to SDS-PAGE. ab85550 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-PMP70 antibody (ab85550) at 1 µg/mL

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

ABCD3 (PMP70) knockout HAP1 whole cell lysate at 20 µg

Lane 3:

A431 whole cell lysate at 20 µg

Lane 4:

HepG2 whole cell lysate at 20 µg

Predicted band size: 75 kDa

false

• WB

Unknown

Western blot - Anti-PMP70 antibody (AB85550)

All lanes:

Western blot - Anti-PMP70 antibody (ab85550) at 1 µg/mL

All lanes:

Lung (Mouse) 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: 75 kDa

Observed band size: 70 kDa

true

Exposure time: 1min