Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab6721)

ab8895 is specific for mono-methylated Lysine 4 of histone H3 and does not recognize di- or tri-methyl Lysine 4 nor methylation at Lysine 9. This is shown in lane 2 where the activity of the antibody is specifically blocked by the addition of the immunizing peptide (ab1340).

All lanes:

Western blot - Anti-Histone H3 (mono methyl K4) antibody - ChIP Grade (ab8895) at 1/500 dilution

All lanes:

Calf thymus histone lysate

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution

Performed under reducing conditions.

Predicted band size: 15 kDa

Observed band size: 18 kDa

Exposure time: 2min

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab6721)

ab9053 is specific for Histone H4 (tri-methyl K20). This is illustrated in lane 5 where the activity of ab9053 is specifically blocked by the addition of the immunizing peptide (ab17567).

All lanes:

Western blot - Anti-Histone H4 (tri methyl K20) antibody - ChIP Grade (ab9053) at 1 µg/mL

All lanes:

Calf thymus histone lysate

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution

Predicted band size: 11 kDa

Observed band size: 13 kDa

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab6721)

Rabbit polyclonal to RAD54 (ab10705) at 1/500.

Lane 1 Marker

Lane 2 HeLa nuclear extract

Lane 3 HeLa cell lysate

Lane 4 A432 cell lysate

Lane 5 MCF7 cell lysate

Lane 6 HEK 293 cell lysate

Secondary antibody - Goat polyclonal to rabbit IgG (HRP) - ab6721.

Lane 1:

Western blot - Anti-RAD54 antibody (ab10705)

Lanes 2 - 6:

Western blot - Anti-RAD54 antibody (ab10705) at 1/500 dilution

Lane 1:

Marker

Lane 2:

HeLa nuclear cell lysate

Lane 3:

HeLa cell lysate

Lane 4:

A432 cell lysate

Lane 5:

MCF7 cell lysate

Lane 6:

Hek 293 cell lysate

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab6721)

Predicted band size: 84 kDa

Observed band size: 70 kDa

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab6721)

All lanes:

Western blot - Anti-Histone H2A.Z antibody - ChIP Grade (ab4174) at 1/500 dilution

All lanes:

Calf thymus histone lysate

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution

Performed under reducing conditions.

Predicted band size: 14 kDa

Observed band size: 15 kDa

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab6721)

Lysates:

Lane 1,3,5: 0.5µg/lane untreated histone

Lane 2,4,6: 0.5µg/lane colcemid treated histone - treated

Primary antibody:

Lane 1-2 : ab5176 (1g/ml)

Lane 3-4 :ab5176 (1g/ml) + phospho peptide (ab5176) 1g/ml

Lane 5-6 :ab5176 (1g/ml) + non-phospho peptide ( ab2903 batch 3) 1g/ml

All lanes:

Western blot - Anti-Histone H3 (phospho S10) antibody (ab5176) at 1 µg/mL

Lanes 1, 3 and 5:

untreated histones at 0.5 µg

Lanes 2, 4 and 6:

colcemid treated histones at 0.5 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 15 kDa

Observed band size: 15-20 kDa

Exposure time: 10s