Anti-PCK1/PEPC antibody [EPR6938] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PCK1/PEPC antibody [EPR6938] - BSA and Azide free (AB248573)

This data was developed using ab133603, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human liver tissue labelling PCK1/PEPC with ab133603 at 1/100.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-PCK1/PEPC antibody [EPR6938] - BSA and Azide free (AB248573)

This data was developed using the same antibody clone in a different buffer formulation (ab133603).

Western blot : Anti-PCK1 antibody [EPR6938] (ab133603) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab133603 was shown to bind specifically to PCK1. A band was observed at 67 kDa in wild-type A549 cell lysates with no signal observed at this size in PCK1 knockout cell line. To generate this image, wild-type and PCK1 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % 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 IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-PCK1/PEPC antibody [EPR6938] (<a href='/en-us/products/primary-antibodies/pck1-pepc-antibody-epr6938-ab133603'>ab133603</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

PCK1 knockout A549 cell lysate at 20 µg

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

HEK-293 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 67 kDa

false

• WB

Unknown

Western blot - Anti-PCK1/PEPC antibody [EPR6938] - BSA and Azide free (AB248573)

This data was developed using ab133603, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-PCK1/PEPC antibody [EPR6938] (<a href='/en-us/products/primary-antibodies/pck1-pepc-antibody-epr6938-ab133603'>ab133603</a>) at 1/10000 dilution

Lane 1:

Human fetal kidney tissue lysate

Lane 2:

Human adipose tissue lysate

Secondary

All lanes:

Goat anti-Rabbit HRP at 1/2000 dilution

Predicted band size: 39 kDa,69 kDa

Observed band size: 39 kDa,69 kDa

false

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-PCK1/PEPC antibody [EPR6938] - BSA and Azide free (AB248573)

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.