Anti-APPL antibody [EPR13569]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APPL antibody [EPR13569] (AB180140)

Immunohistochemical analysis of paraffin-embedded Human colon carcinoma tissue labeling APPL with ab180140 at 1/100 dilution. Secondary antibody Goat anti rabbit IgG(HRP) prediluted. Counter stain Hematoxylin.

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-APPL antibody [EPR13569] (AB180140)

Immunofluorescent analysis of 4% paraformaldehyde fixed MCF7 cells labeling APPL with ab180140 at 1/500 dilution. Secondary antibody Goat anti rabbit IgG(Dylight 555) Conterstained with Dapi blue.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APPL antibody [EPR13569] (AB180140)

Immunohistochemical analysis of paraffin-embedded Human pancreas carcinoma tissue labeling APPL with ab180140 at 1/100 dilution. Secondary antibody Goat anti rabbit IgG(HRP) prediluted. Counter stain Hematoxylin.

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-APPL antibody [EPR13569] (AB180140)

Lanes 1- 4 : Merged signal (red and green). Green - ab180140 observed at 90 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab180140 was shown to react with Anti-APPL in wild-type HeLa cells in western blot. The band observed in knockout cell line ab265187 (knockout cell lysate ab257836) lane below 90kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and APPL1 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. ab180140 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 10000 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-APPL antibody [EPR13569] (ab180140) at 1/10000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

APPL knockout HeLa cell lysate at 20 µg

Lane 3:

MCF7 cell lysate at 20 µg

Lane 4:

HEK-293T cell lysate at 20 µg

Predicted band size: 80 kDa

Observed band size: 90 kDa

false

• WB

Supplier Data

Western blot - Anti-APPL antibody [EPR13569] (AB180140)

All lanes:

Western blot - Anti-APPL antibody [EPR13569] (ab180140) at 1/10000 dilution

Lane 1:

HeLa cell lysate at 20 µg

Lane 2:

MCF7 cell lysate at 20 µg

Lane 3:

293 cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 80 kDa

Observed band size: 90 kDa

false

• WB

Lab

Western blot - Anti-APPL antibody [EPR13569] (AB180140)

Lanes 1- 4 : Merged signal (red and green). Green - ab180140 observed at 90 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab180140 was shown to react with Anti-APPL in wild-type HeLa cells in western blot. The band observed in CRISPR/Cas9 edited cell line ab265187 (CRISPR/Cas9 edited cell lysate ab257836) lane below 90kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and APPL1 CRISPR/Cas9 edited 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. ab180140 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 10000 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-APPL antibody [EPR13569] (ab180140) at 1/10000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

APPL CRISPR/Cas9 edited HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human APPL1 (APPL) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-appl1-appl-knockout-hela-cell-line-ab265187'>ab265187</a>)

Lane 3:

MCF7 cell lysate at 20 µg

Lane 4:

HEK-293T cell lysate at 20 µg

Predicted band size: 80 kDa

Observed band size: 90 kDa

false