Anti-APPL antibody [EPR13569] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APPL antibody [EPR13569] - BSA and Azide free (AB250150)

This data was developed using ab180140, the same antibody clone in a different buffer formulation.

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] - BSA and Azide free (AB250150)

This data was developed using ab180140, the same antibody clone in a different buffer formulation.

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] - BSA and Azide free (AB250150)

This data was developed using ab180140, the same antibody clone in a different buffer formulation.

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

Supplier Data

Western blot - Anti-APPL antibody [EPR13569] - BSA and Azide free (AB250150)

This data was developed using ab180140, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-APPL antibody [EPR13569] (<a href='/en-us/products/primary-antibodies/appl-antibody-epr13569-ab180140'>ab180140</a>) 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] - BSA and Azide free (AB250150)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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] (<a href='/en-us/products/primary-antibodies/appl-antibody-epr13569-ab180140'>ab180140</a>) 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