Anti-Amyloid Precursor Protein antibody [EPR5119(2)] - BSA and Azide free

• IP

Unknown

Immunoprecipitation - Anti-Amyloid Precursor Protein antibody [EPR5119(2)] - BSA and Azide free (AB248567)

This data was developed using ab133588, the same antibody clone in a different buffer formulation.

ab133588 (purified) at 1/20 immunoprecipitating amyloid precursor protein in human fetal brain (Lane 2). Lane 3 - PBS. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).

Blocking buffer and concentration : 5% NFDM/TBST.

Diluting buffer and concentration : 5% NFDM /TBST.

All lanes:

Immunoprecipitation - Anti-Amyloid Precursor Protein antibody [EPR5119(2)] (<a href='/en-us/products/primary-antibodies/amyloid-precursor-protein-antibody-epr51192-ab133588'>ab133588</a>)

Predicted band size: 86 kDa

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• WB

Unknown

Western blot - Anti-Amyloid Precursor Protein antibody [EPR5119(2)] - BSA and Azide free (AB248567)

This data was developed using ab133588, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Amyloid Precursor Protein antibody [EPR5119(2)] (<a href='/en-us/products/primary-antibodies/amyloid-precursor-protein-antibody-epr51192-ab133588'>ab133588</a>) at 1/2000 dilution

All lanes:

HEK-293 cell lysate at 20 µg

Secondary

All lanes:

HRP conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 148 kDa,86 kDa

Observed band size: 152 kDa

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• WB

Lab

Western blot - Anti-Amyloid Precursor Protein antibody [EPR5119(2)] - BSA and Azide free (AB248567)

This data was developed using ab133588, the same antibody clone in a different buffer formulation.

Lane 1 : Wild type HAP1 whole cell lysate (20 μg)

Lane 2 : Amyloid Precursor Protein 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 - ab133588 observed at 110 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab133588 was shown to recognize Amyloid Precursor Protein when Amyloid Precursor Protein knockout samples were used, along with additional cross-reactive bands. Wild-type and Amyloid Precursor Protein knockout samples were subjected to SDS-PAGE. ab133588 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 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.

All lanes:

Western blot - Anti-Amyloid Precursor Protein antibody [EPR5119(2)] (<a href='/en-us/products/primary-antibodies/amyloid-precursor-protein-antibody-epr51192-ab133588'>ab133588</a>)

Predicted band size: 86 kDa

false

• WB

Unknown

Western blot - Anti-Amyloid Precursor Protein antibody [EPR5119(2)] - BSA and Azide free (AB248567)

This data was developed using ab133588, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Amyloid Precursor Protein antibody [EPR5119(2)] (<a href='/en-us/products/primary-antibodies/amyloid-precursor-protein-antibody-epr51192-ab133588'>ab133588</a>) at 1/2000 dilution

All lanes:

SH-SY5Y cell lysate at 20 µg

Secondary

All lanes:

HRP conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 13 kDa,86 kDa

false

• WB

Unknown

Western blot - Anti-Amyloid Precursor Protein antibody [EPR5119(2)] - BSA and Azide free (AB248567)

This data was developed using ab133588, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Amyloid Precursor Protein antibody [EPR5119(2)] (<a href='/en-us/products/primary-antibodies/amyloid-precursor-protein-antibody-epr51192-ab133588'>ab133588</a>) at 1/20000 dilution

All lanes:

Human fetal brain tissue lysate

Secondary

All lanes:

HRP conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 86 kDa

false

• WB

Unknown

Western blot - Anti-Amyloid Precursor Protein antibody [EPR5119(2)] - BSA and Azide free (AB248567)

This data was developed using ab133588, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-Amyloid Precursor Protein antibody [EPR5119(2)] (<a href='/en-us/products/primary-antibodies/amyloid-precursor-protein-antibody-epr51192-ab133588'>ab133588</a>) at 1/1000 dilution

Lane 1:

293T cell lysate at 10 µg

Lane 2:

SH-SY5Y cell lysate at 10 µg

Lane 3:

U87 MG cell lysate at 10 µg

Lane 4:

Human fetal brain tissue lysate at 10 µg

Secondary

All lanes:

HRP labelled Goat anti-Rabbit IgG at 1/2000 dilution

Predicted band size: 86 kDa

Observed band size: 100-120 kDa

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• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-Amyloid Precursor Protein antibody [EPR5119(2)] - BSA and Azide free (AB248567)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.