Anti-EZH2 antibody [EPR24902-112]

• WB

Supplier Data

Western blot - Anti-EZH2 antibody [EPR24902-112] (AB287893)

ab287893 was shown to specifically react with EZH2 (pT345) in untreated Wild-type HAP1, HeLa and NIH/3T3 (Figure A). After treatment with 150 units alkaline phosphatase for 1 hour at 37°C, no band was observed (Figure B). Samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% BSA in TBS-0.1 % Tween® 20 (TBS-T). ab287893 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 and 1 in 20000 dilutions, respectively. Blots were developed with Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.​

All lanes:

Western blot - Anti-EZH2 antibody [EPR24902-112] (ab287893) at 1/1000 dilution

Lane 1:

Wild-type HAP1 cell lysate at 20 µg

Lane 2:

EZH2 knockout HAP1 cell lysate​ at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

NIH/3T3 cell lysate at 20 µg

Secondary

Lanes 1 - 4:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 4:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 98 kDa,37 kDa

false

• WB

Supplier Data

Western blot - Anti-EZH2 antibody [EPR24902-112] (AB287893)

ab287893 was shown to specifically react with EZH2 (pT345) in HEK-293. The signal was significantly reduced after blocking with 1µg/ml histone-lysine N-methyltransferase EZH2 peptide (pT345). Samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% BSA in TBS-0.1 % Tween® 20 (TBS-T). ab287893 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 and 1 in 20000 dilutions, respectively. Blots were developed with Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.​

All lanes:

Western blot - Anti-EZH2 antibody [EPR24902-112] (ab287893) at 1/1000 dilution

Lane 1:

HEK-293 cell lysate at 20 µg

Lane 2:

HEK-293 cell lysate with histone-lysine N-methyltransferase EZH2 peptide (unmodified) at 1µg/ml​ at 20 µg

Lane 3:

HEK-293 cell lysate with histone-lysine N-methyltransferase EZH2 peptide (pT345) at 1µg/ml at 20 µg

Secondary

Lanes 1 - 3:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 3:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 98 kDa,37 kDa

false

• WB

Supplier Data

Western blot - Anti-EZH2 antibody [EPR24902-112] (AB287893)

ab287893 was shown to specifically react with EZH2 (pT345) in HeLa and NIH/3T3. Samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% BSA in TBS-0.1 % Tween® 20 (TBS-T). ab287893 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 and 1 in 20000 dilutions, respectively. Blots were developed with Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.​

All lanes:

Western blot - Anti-EZH2 antibody [EPR24902-112] (ab287893) at 1/1000 dilution

Lane 1:

HeLa cell lysate at 20 µg

Lane 2:

NIH/3T3 cell lysate​ at 20 µg

Secondary

Lanes 1 - 2:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 2:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 98 kDa,37 kDa

false

• I-ELISA

Supplier Data

Indirect ELISA - Anti-EZH2 antibody [EPR24902-112] (AB287893)

Indirect ELISA showing the phospho-specificity (pT345) of primary antibody ab287893. Plates were coated with histone-lysine N-methyltransferase EZH2 (pT345), histone-lysine N-methyltransferase EZH2 (pT345), histone-lysine N-methyltransferase EZH2 peptide (unmodified) and histone-lysine N-methyltransferase EZH2 peptide (pT487) peptides at 1000 ng/ml. Binding of ab286909 was assessed in a serial dilution range 0.016- 1000 ng/mL (a 3-fold serial dilution).​

Binding was detected using pre-adsorbed secondary antibody, goat anti-rabbit IgG H&L (HRP, ab97080) at 1/2000 dilution.​