Anti-ATP7b antibody [EPR6794]

• WB

Lab

Western blot - Anti-ATP7b antibody [EPR6794] (AB124973)

Western blot : Rabbit Monoclonal[EPR6794] to ATP7b ab124973 staining at 1/1000 dilution, shown in green; Mouse anti-CANX (ab238078) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 153 kDa in Wild-type A549 ab288558 cell lysates with no signal observed at this size in ATP7B knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-ATP7b antibody [EPR6794] (ab124973) at 1/1000 dilution

Lane 1:

Wild-type A549 at 10 µg

Lane 2:

ATP7B knockout A549 at 10 µg

Lane 3:

HepG2 at 10 µg

Lane 4:

Ramos at 10 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 157 kDa

Observed band size: 153 kDa

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• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-ATP7b antibody [EPR6794] (AB124973)

Intracellular Flow Cytometry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling ATP7b with purified ab124973 at 1/20 dilution (10μg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluorr^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-ATP7b antibody [EPR6794] (AB124973)

Immunocytochemistry/ Immunofluorescence analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling ATP7b with purified ab124973 at 1 : 100 dilution (1.2 μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1 : 200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1 : 1000 (2 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-ATP7b antibody [EPR6794] (AB124973)

Intracellular flow cytometric analysis of permeabilized HepG2 cells using unpurified anti-ATP7B RabMAb in red (ab124973) or a rabbit IgG in green (negative).

• WB

Lab

Western blot - Anti-ATP7b antibody [EPR6794] (AB124973)

All lanes:

Western blot - Anti-ATP7b antibody [EPR6794] (ab124973) at 1/1000 dilution

All lanes:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 157 kDa

Observed band size: 157 kDa

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• WB

Lab

Western blot - Anti-ATP7b antibody [EPR6794] (AB124973)

All lanes:

Western blot - Anti-ATP7b antibody [EPR6794] (ab124973) at 1/10000 dilution

All lanes:

Caco-2 (Human colorectal adenocarcinoma epithelial cell) whole cell lysates at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 157 kDa

Observed band size: 157 kDa

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• OI-RD Scanning

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OI-RD Scanning - Anti-ATP7b antibody [EPR6794] (AB124973)

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.