Anti-Amyloid Precursor Protein antibody [EPR5118-34] - BSA and Azide free

• WB

Lab

Western blot - Anti-Amyloid Precursor Protein antibody [EPR5118-34] - BSA and Azide free (AB238916)

ab126732 was shown to react with APP in wild-type HAP1 cells in Western blot with loss of signal observed in a APP knockout cell line. Wild-type HAP1 and APP knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab126732 overnight at 4 °C at a 1/500 dilution. Blots were incubated with secondary antibodies at 0.2ug/mL before imaging.

These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Western blot - Anti-Amyloid Precursor Protein antibody [EPR5118-34] (<a href='/en-us/products/primary-antibodies/amyloid-precursor-protein-antibody-epr5118-34-ab126732'>ab126732</a>) at 1/500 dilution

Lane 1:

Wild-type HAP1 lysate at 20 µg

Lane 2:

APP Knockout HAP1 lysate at 20 µg

Observed band size: 100 kDa

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• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Amyloid Precursor Protein antibody [EPR5118-34] - BSA and Azide free (AB238916)

ab126732, at a dilution of 1/100, staining Amyloid Precursor Protein in paraffin-embedded Human fetal brain tissue by Immunohistochemistry.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab126732).

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Amyloid Precursor Protein antibody [EPR5118-34] - BSA and Azide free (AB238916)

Overlay histogram showing A431 cells stained with ab126732 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab126732, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab126732).

• WB

Supplier Data

Western blot - Anti-Amyloid Precursor Protein antibody [EPR5118-34] - BSA and Azide free (AB238916)

Lane 1 : Wild type HAP1 whole cell lysate (20 μg)
Lane 2 : APP knockout HAP1 whole cell lysate (20 μg)
Lane 3 : HepG2 whole cell lysate (20 μg)
Lane 4 : HeLa whole cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab126732 observed at 110 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab126732 was shown to specifically react with APP when APP knockout samples were used. Wild-type and APP knockout samples were subjected to SDS-PAGE. ab126732 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1000 dilution and 1/10000 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 1/10000 dilution for 1 hour at room temperature before imaging.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab126732).

All lanes:

Western blot - Anti-Amyloid Precursor Protein antibody [EPR5118-34] - BSA and Azide free (ab238916)

Predicted band size: 86 kDa

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