Anti-GATA1 antibody [EPR2820(2)]

• WB

Unknown

Western blot - Anti-GATA1 antibody [EPR2820(2)] (AB133274)

All lanes:

Western blot - Anti-GATA1 antibody [EPR2820(2)] (ab133274) at 1/1000 dilution

Lane 1:

Molt-4 cell lysate

Lane 2:

TF-1 cell lysate

Lane 3:

K562 cell lysate

Lane 4:

HL-60 cell lysate

Predicted band size: 43 kDa

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• WB

Lab

Western blot - Anti-GATA1 antibody [EPR2820(2)] (AB133274)

False colour image of Western blot : Anti-GATA1 antibody [EPR2820(2)] staining at 1/10000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab133274 was shown to bind specifically to GATA1. A band was observed at 47 kDa in wild-type K562 cell lysates with no signal observed at this size in GATA1 knockout cell line. To generate this image, wild-type and GATA1 knockout K562 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-GATA1 antibody [EPR2820(2)] (ab133274) at 1/10000 dilution

Lane 1:

Wild-type K562 cell lysate at 20 µg

Lane 2:

GATA1 [C116] knockout K562 cell lysate at 20 µg

Lane 3:

MOLT-4 cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Observed band size: 47 kDa

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• WB

Lab

Western blot - Anti-GATA1 antibody [EPR2820(2)] (AB133274)

False colour image of Western blot : Anti-GATA1 antibody [EPR2820(2)] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab133274 was shown to bind specifically to GATA1. A band was observed at 47 kDa in wild-type K562 cell lysates with no signal observed at this size in GATA1 knockout cell line ab285360 (knockout cell lysate ab289686). To generate this image, wild-type and GATA1 knockout K562 cell lysates were analysed. 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-GATA1 antibody [EPR2820(2)] (ab133274) at 1/1000 dilution

Lane 1:

Wild-type K562 cell lysate at 20 µg

Lane 2:

GATA1 knockout K562 cell lysate at 20 µg

Lane 3:

HL-60 cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Observed band size: 47 kDa

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• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-GATA1 antibody [EPR2820(2)] (AB133274)

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.