Anti-TUG antibody [EPR8615]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TUG antibody [EPR8615] (AB131237)

ab131237 at 1/100 staining TUG in Formalin-fixed Paraffin-embedded Human colon tissue by Immunohistochemistry.

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

• WB

Lab

Western blot - Anti-TUG antibody [EPR8615] (AB131237)

Western blot : Anti-TUG antibody [EPR8615] ab131237 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 70 kDa in Wild-type A549 cell lysates with no signal observed at this size in ASPSCR1 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 3pc 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 and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-TUG antibody [EPR8615] (ab131237) at 1/1000 dilution

Lane 1:

Wild-type A549 whole cell lysate at 20 µg

Lane 2:

ASPSCR1 knockout A549 whole cell lysate at 20 µg

Lane 3:

HEK-293 whole cell lysate at 20 µg

Lane 4:

HeLa whole cell lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 70 kDa

Observed band size: 70 kDa

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• WB

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Western blot - Anti-TUG antibody [EPR8615] (AB131237)

All lanes:

Western blot - Anti-TUG antibody [EPR8615] (ab131237) at 1/10000 dilution

Lane 1:

K562 cell lysate at 10 µg

Lane 2:

293T cell lysate at 10 µg

Lane 3:

SW480 cell lysate at 10 µg

Predicted band size: 60 kDa

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• WB

Lab

Western blot - Anti-TUG antibody [EPR8615] (AB131237)

Lane 1 : Wild-type HAP1 cell lysate (20 μg)
Lane 2 : TUG knockout HAP1 cell lysate (20 μg)
Lane 3 : NIH/3T3 cell lysate (20 μg)
Lane 4 : HEK293 cell lysate (20 μg)
Lanes 1 - 4 : Merged signal (red and green). Green - ab131237 observed at 70 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab131237 was shown to specifically react with TUG when TUG knockout samples were used. Wild-type and TUG knockout samples were subjected to SDS-PAGE. ab131237 and ab8245 (loading control to GAPDH) were both diluted 1/10 000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-TUG antibody [EPR8615] (ab131237)

Predicted band size: 60 kDa,69 kDa

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• OI-RD Scanning

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OI-RD Scanning - Anti-TUG antibody [EPR8615] (AB131237)

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.