Anti-PMP70 antibody [EPR5614] - BSA and Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-PMP70 antibody [EPR5614] - BSA and Azide free (AB247868)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling PMP70 with primary antibody anti-PMP70 (ab109448) at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) (ab150081) secondary antibody at 1/1000 dilution (2.0 μg/mL). Confocal image showing cytoplasmic staining on HeLa cells. Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) (ab195889) was used to counterstain tubulin at 1/200 dilution (2.5 μg/mL). The nuclear counter stain is DAPI (blue).
The secondary antibody only control is : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) at 1/1000 dilution (2.0 μg/mL).

This data was developed using the same antibody clone in a different buffer formulation (ab109448).

• WB

Lab

Western blot - Anti-PMP70 antibody [EPR5614] - BSA and Azide free (AB247868)

This data was developed using the same antibody clone in a different buffer formulation (ab109448).

Lanes 1 - 4 : Merged signal (red and green). Green - ab109448 observed at 75 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab109448 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. ab109448 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution 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 [EPR5614] - BSA and Azide free (ab247868) at 1/1000 dilution

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

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• WB

Lab

Western blot - Anti-PMP70 antibody [EPR5614] - BSA and Azide free (AB247868)

This data was developed using the same antibody clone in a different buffer formulation (ab109448).

Lanes 1- 2 : Merged signal (red and green). Green - ab109448 observed at 75 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab109448 was shown to react with PMP70 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265294 (knockout cell lysate ab257134) was used. Wild-type HeLa and PMP70 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109448 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-PMP70 antibody [EPR5614] (<a href='/en-us/products/primary-antibodies/pmp70-antibody-epr5614-ab109448'>ab109448</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 40 µg

Lane 2:

ABCD3 knockout HeLa cell lysate at 40 µg

Predicted band size: 75 kDa

Observed band size: 75 kDa

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