Human Histone H3 (unmodified) peptide

• WB

Ap

Western blot - Human Histone H3 (unmodified) peptide (AB7228)

This data was developed using the same antibody clone in a different formulation containing PBS and Azide (ab14955).

All lanes:

Western blot - Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] (<a href='/en-us/products/primary-antibodies/histone-h3-phospho-s10-antibody-mabcam-14955-ab14955'>ab14955</a>) at 1 µg/mL

Lanes 1, 5 and 7:

Control HeLa Histone Prep at 0.5 µg

Lanes 2, 6 and 8:

Colcemid treated HeLa Histone prep at 0.5 µg

Lane 3:

Control HeLa Histone Prep at 0.5 µg with Human Histone H3 (tri methyl K9, phospho S10) peptide (<a href='/en-us/products/unavailable/human-histone-h3-tri-methyl-k9-phospho-s10-peptide-ab15644'>ab15644</a>)

Lane 4:

Colcemid treated HeLa Histone prep at 0.5 µg with Human Histone H3 (tri methyl K9, phospho S10) peptide (<a href='/en-us/products/unavailable/human-histone-h3-tri-methyl-k9-phospho-s10-peptide-ab15644'>ab15644</a>)

Lane 9:

Control HeLa Histone Prep at 0.5 µg with Human Histone H3 (phospho S28) peptide (<a href='/en-us/products/unavailable/human-histone-h3-phospho-s28-peptide-ab14793'>ab14793</a>)

Lane 10:

Colcemid treated HeLa Histone prep at 0.5 µg with Human Histone H3 (phospho S28) peptide (<a href='/en-us/products/unavailable/human-histone-h3-phospho-s28-peptide-ab14793'>ab14793</a>)

Secondary

All lanes:

Rabbit polyclonal to Mouse IgG H&L (HRP) at 1/5000 dilution

Predicted band size: 15 kDa

Observed band size: 17 kDa

false

• WB

Unknown

Western blot - Human Histone H3 (unmodified) peptide (AB7228)

ab5819 is specific for Histone H3 phosphorylated at residue S10 and tri methylated at residue K9. The activity of the antibody is blocked by the addition of the immunizing peptide, ab15644 (lanes 3 and 4).

All lanes:

Western blot - Anti-Histone H3 (tri methyl K9, phospho S10) antibody (<a href='/en-us/products/primary-antibodies/histone-h3-tri-methyl-k9-phospho-s10-antibody-ab5819'>ab5819</a>) at 1/500 dilution

Lanes 1, 5, 7 and 9:

HeLa histones at 0.5 µg

Lanes 2, 6, 8 and 10:

Colcemid-treated HeLa histones at 0.5 µg

Lane 3:

HeLa histones at 0.5 µg with Human Histone H3 (tri methyl K9, phospho S10) peptide (<a href='/en-us/products/unavailable/human-histone-h3-tri-methyl-k9-phospho-s10-peptide-ab15644'>ab15644</a>)

Lane 4:

Colcemid-treated HeLa histones at 0.5 µg with Human Histone H3 (tri methyl K9, phospho S10) peptide (<a href='/en-us/products/unavailable/human-histone-h3-tri-methyl-k9-phospho-s10-peptide-ab15644'>ab15644</a>)

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab6721'>ab6721</a>) at 1/5000 dilution

Predicted band size: 15 kDa

Observed band size: 22 kDa,60 kDa

false

• WB

Project

Western blot - Human Histone H3 (unmodified) peptide (AB7228)

This data was developed using the same antibody clone in a different formulation containing PBS and Azide (ab14955).

All lanes:

Western blot - Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] (<a href='/en-us/products/primary-antibodies/histone-h3-phospho-s10-antibody-mabcam-14955-ab14955'>ab14955</a>) at 0.5 µg/mL

Lanes 1, 3 and 7:

Control HeLa Histone prep at 5 µg

Lanes 2, 4 and 8:

Colecemid treated HeLa Histone prep at 5 µg

Lane 5:

Control HeLa Histone prep at 5 µg with Human Histone H3 (phospho S28) peptide (<a href='/en-us/products/unavailable/human-histone-h3-phospho-s28-peptide-ab14793'>ab14793</a>)

Lane 6:

Colecemid treated HeLa Histone prep at 5 µg with Human Histone H3 (phospho S28) peptide (<a href='/en-us/products/unavailable/human-histone-h3-phospho-s28-peptide-ab14793'>ab14793</a>)

Secondary

All lanes:

Western blot - Rabbit Anti-Mouse IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/rabbit-mouse-igg-h-l-hrp-ab6728'>ab6728</a>) at 1/5000 dilution

Predicted band size: 15 kDa

Observed band size: 20 kDa

false

• WB

Project1592****

Western blot - Human Histone H3 (unmodified) peptide (AB7228)

Expected molecular weights : H3 = 17 kDa; H4 = 14 kDa This image shows that the main epitopes recognized by ab7315 are the di methylated lysine residues. This can be seen in lanes 4, 7 and 10 where the activity of the antibody is quenched by the immunizing peptides (ab7768, ab1772, ab1781).

All lanes:

Western blot - Anti-pan methyl Lysine antibody - ChIP Grade (<a href='/en-us/products/primary-antibodies/pan-methyl-lysine-antibody-chip-grade-ab7315'>ab7315</a>) at 1 µg/mL

All lanes:

Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 14-17 kDa

true

Exposure time: 20min

• WB

Lab

Western blot - Human Histone H3 (unmodified) peptide (AB7228)

Lane 8 shows that Rabbit polyclonal to Histone H3 (tri methyl K9) is blocked by the addition of the immunizing peptide (ab1773). Cross-reactivity with Histone H3 peptide - tri methyl K27 (ab1782) is also shown in Lane 11.

All lanes:

Western blot - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (<a href='/en-us/products/primary-antibodies/histone-h3-tri-methyl-k9-antibody-chip-grade-ab8898'>ab8898</a>) at 1 µg/mL

All lanes:

Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg

Secondary

All lanes:

IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Predicted band size: 15 kDa

Observed band size: 17 kDa

false

• WB

Project8270****

Western blot - Human Histone H3 (unmodified) peptide (AB7228)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine, and sodium azide (ab12209).

All lanes:

Western blot - Anti-Histone H3 (tri methyl K4) antibody [mAbcam12209] - ChIP Grade (<a href='/en-us/products/primary-antibodies/histone-h3-tri-methyl-k4-antibody-mabcam12209-chip-grade-ab12209'>ab12209</a>) at 2 µg/mL

All lanes:

Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg

Secondary

All lanes:

Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 15 kDa

Observed band size: 17 kDa

false

Exposure time: 16min

• WB

Project

Western blot - Human Histone H3 (unmodified) peptide (AB7228)

All lanes:

Western blot - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (<a href='/en-us/products/primary-antibodies/histone-h3-mono-methyl-k9-antibody-chip-grade-ab9045'>ab9045</a>) at 1 µg/mL

All lanes:

Calf Thymus Histone Preparation Nuclear Lysate (<a href='/en-us/products/unavailable/calf-thymus-histone-preparation-nuclear-lysate-ab121'>ab121</a>) at 0.5 µg

Secondary

All lanes:

IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Predicted band size: 15 kDa

false

• WB

Unknown

Western blot - Human Histone H3 (unmodified) peptide (AB7228)

Rabbit polyclonal to Histone H3 K9 Methyl K9 (1/1000)

Peptides at 1 ug/ml

1XTBS, 5%BSA, 0.5% Tween

This antibody shows significantly greater reactivity with mono methyl K9. This can be seen in lane 7, as the addition of ab1771 (mono methyl K9) completely blocks the activity of ab9045. Weaker cross-reactivity is seen against mono methyl K27. This is shown in lane 3, as the addition of ab1780 only partially blocks the activity of ab9045.

All lanes:

Western blot - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (<a href='/en-us/products/primary-antibodies/histone-h3-mono-methyl-k9-antibody-chip-grade-ab9045'>ab9045</a>) at 1 µg/mL

Predicted band size: 15 kDa

false

• WB

Project

Western blot - Human Histone H3 (unmodified) peptide (AB7228)

ab8896 specifically recognises mono-methyl K9 (lane 1) in calf thymus lysate at 17kDa. ab8896 is successfully blocked using the immunizing peptide (lane 2 ab1771), but not the di-methyl K9 (lane 3 ab1772), the tri-methyl K9 (lane 4 ab1773), the mono-methyl K27 (lane 5 ab1780), the di-methyl K27 (lane 6 ab1781), the tri-methyl K27 (lane 7 ab1782), the mono-methyl K4 (lane 8 ab1340) nor the corresponding unmodified peptides (lane 9 ab7228, lane 10 ab2623). This implies that ab8896 is specific to mono-methyl K9.

All lanes:

Western blot - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (<a href='/en-us/products/primary-antibodies/histone-h3-mono-methyl-k9-antibody-chip-grade-ab8896'>ab8896</a>) at 1 µg/mL

All lanes:

Calf Thymus Histone Preparation at 0.5 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG H&L (HRP) Pre-Adsorbed at 1/10000 dilution

Predicted band size: 15 kDa

Observed band size: 17 kDa

false

• WB

Unknown

Western blot - Human Histone H3 (unmodified) peptide (AB7228)

All lanes:

Western blot - Anti-Histone H3 (di methyl K4, tri methyl K4) antibody [mAbcam 6000] - ChIP Grade (<a href='/en-us/products/primary-antibodies/histone-h3-di-methyl-k4-tri-methyl-k4-antibody-mabcam-6000-chip-grade-ab6000'>ab6000</a>) at 1 µg/mL

All lanes:

Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-hrp-preadsorbed-ab97040'>ab97040</a>) at 1/10000 dilution

Predicted band size: 15 kDa

Observed band size: 17 kDa

true

Exposure time: 20min