Anti-PGAM1 antibody [EPR8042]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-PGAM1 antibody [EPR8042] (AB129191)

ab129191 at 1/100 dilution staining PGAM1 in permeabilized Raji cells by intracellular flow cytometry or a rabbit IgG negative control (green).

• WB

Lab

Western blot - Anti-PGAM1 antibody [EPR8042] (AB129191)

Western blot : Anti-PGAM antibody [EPR8042] (ab129191) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab129191 was shown to bind specifically to PGAM. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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-PGAM1 antibody [EPR8042] (ab129191) at 1/1000 dilution

Lane 1:

Recombinant human PGAM2 protein (Active) <a href='/en-us/products/proteins-peptides/recombinant-human-pgam2-protein-active-ab219276'>ab219276</a> cell lysate at 0.1 µg

Lane 2:

Recombinant human PGAM1 protein (Active) <a href='/en-us/products/proteins-peptides/recombinant-human-pgam1-protein-active-ab289791'>ab289791</a> cell lysate at 0.1 µg

Lane 3:

PGAM4 Human Recombinant Protein cell lysate at 0.1 µg

Lane 4:

Jurkat cell lysate at 20 µg

Lane 5:

A431 cell lysate at 20 µg

Lane 6:

Raji 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

Predicted band size: 29 kDa

Observed band size: 29 kDa

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• WB

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Western blot - Anti-PGAM1 antibody [EPR8042] (AB129191)

All lanes:

Western blot - Anti-PGAM1 antibody [EPR8042] (ab129191) at 1/1000 dilution

Lane 1:

Jurkat cell lysate at 10 µg

Lane 2:

Human fetal muscle tissue lysate at 10 µg

Lane 3:

A431 cell lysate at 10 µg

Lane 4:

Raji cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 29 kDa

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• OI-RD Scanning

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OI-RD Scanning - Anti-PGAM1 antibody [EPR8042] (AB129191)

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.