Human Histone H3 (tri methyl K4) peptide

• ChIP

Project7459****

ChIP - Human Histone H3 (tri methyl K4) peptide (AB1342)

Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 5µg of ab78513 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.

• WB

Project5783****

Western blot - Human Histone H3 (tri methyl K4) peptide (AB1342)

All lanes:

Western blot - Anti-Histone H4 (mono methyl K20) antibody - ChIP Grade (<a href='/en-us/products/primary-antibodies/histone-h4-mono-methyl-k20-antibody-chip-grade-ab9051'>ab9051</a>) at 1 µg/mL

Lanes 1, 3, 4, 5, 8, 9, 10 and 11:

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

Lane 2:

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 with Human Histone H4 peptide (<a href='/en-us/products/unavailable/human-histone-h4-peptide-ab14963'>ab14963</a>)

Lane 6:

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 with Human Histone H4 (mono methyl K20) peptide (<a href='/en-us/products/unavailable/human-histone-h4-mono-methyl-k20-peptide-ab17043'>ab17043</a>)

Lane 7:

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 with Histone H4 peptide (17-24) - di methyl K20

Secondary

All lanes:

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

Predicted band size: 11 kDa

Observed band size: 13 kDa

false

Exposure time: 10s

• WB

Lab

Western blot - Human Histone H3 (tri methyl K4) peptide (AB1342)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-Arginine and sodium azide (ab78517).

ab78517 is specific for Histone H4 (tri-methyl K20). This is illustrated in lane 8 where the activity of ab78517 is specifically blocked by the addition of the immunizing peptide (ab17567).

All lanes:

Western blot - Anti-Histone H4 (di methyl K20, tri methyl K20) antibody [6F8-D9] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/histone-h4-di-methyl-k20-tri-methyl-k20-antibody-6f8-d9-bsa-and-azide-free-ab264528'>ab264528</a>) at 1 µg/mL

Lanes 1, 3, 4, 5, 9, 10 and 11:

Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg

Lane 2:

Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg with Human Histone H4 peptide (<a href='/en-us/products/unavailable/human-histone-h4-peptide-ab14963'>ab14963</a>)

Lane 6:

Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg with Human Histone H4 (mono methyl K20) peptide (<a href='/en-us/products/unavailable/human-histone-h4-mono-methyl-k20-peptide-ab17043'>ab17043</a>)

Lane 7:

Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg with Histone H4 peptide (17-24) - di methyl K20

Secondary

All lanes:

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

Predicted band size: 11 kDa

Observed band size: 13 kDa

true

Exposure time: 90s

• WB

Project8270****

Western blot - Human Histone H3 (tri methyl K4) peptide (AB1342)

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

Ap

Western blot - Human Histone H3 (tri methyl K4) peptide (AB1342)

All lanes:

Western blot - HRP Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] (<a href='/en-us/products/primary-antibodies/hrp-histone-h3-di-methyl-k9-antibody-mabcam-1220-ab62168'>ab62168</a>) at 1/20000 dilution

All lanes:

Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg

Predicted band size: 15 kDa

Observed band size: 17 kDa

false

• WB

Lab

Western blot - Human Histone H3 (tri methyl K4) peptide (AB1342)

This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab6147 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

All lanes:

Western blot - Anti-Histone H3 (di methyl K27, tri methyl K27) antibody [mAbcam 6147] (<a href='/en-us/products/primary-antibodies/histone-h3-di-methyl-k27-tri-methyl-k27-antibody-mabcam-6147-ab6147'>ab6147</a>) at 1 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 1/5000 dilution

Predicted band size: 15 kDa

Observed band size: 17 kDa

true

Exposure time: 1min

• WB

Project7459****

Western blot - Human Histone H3 (tri methyl K4) peptide (AB1342)

ab78513 is blocked by the addition of Histone H4 (mono methyl K20)peptide (ab17043; lane 6). ab78513 is partially blocked by Histone H4 (di methyl K20) peptide (lane 7).

All lanes:

Western blot - Anti-Histone H4 (mono methyl K20) antibody [5E10-D8] - ChIP Grade (<a href='/en-us/products/primary-antibodies/histone-h4-mono-methyl-k20-antibody-5e10-d8-chip-grade-ab78513'>ab78513</a>) at 1 µg/mL

Lanes 1, 3, 4, 5, 8, 9, 10 and 11:

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

Lane 2:

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 with Human Histone H4 peptide (<a href='/en-us/products/unavailable/human-histone-h4-peptide-ab14963'>ab14963</a>)

Lane 6:

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 with Human Histone H4 (mono methyl K20) peptide (<a href='/en-us/products/unavailable/human-histone-h4-mono-methyl-k20-peptide-ab17043'>ab17043</a>)

Lane 7:

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 with Histone H4 peptide (17-24) - di methyl K20

Secondary

All lanes:

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

Predicted band size: 11 kDa

Observed band size: 13 kDa,17 kDa

true

Exposure time: 1min

• WB

Lab

Western blot - Human Histone H3 (tri methyl K4) peptide (AB1342)

ab78517 is specific for Histone H4 (tri-methyl K20). This is illustrated in lane 8 where the activity of ab78517 is specifically blocked by the addition of the immunizing peptide (ab17567).

All lanes:

Western blot - Anti-Histone H4 (di methyl K20, tri methyl K20) antibody [6F8-D9] (<a href='/en-us/products/primary-antibodies/histone-h4-di-methyl-k20-tri-methyl-k20-antibody-6f8-d9-ab78517'>ab78517</a>) at 1 µg/mL

Lanes 1, 3, 4, 5, 8, 9, 10 and 11:

Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg

Lane 2:

Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg with Human Histone H4 peptide (<a href='/en-us/products/unavailable/human-histone-h4-peptide-ab14963'>ab14963</a>)

Lane 6:

Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg with Human Histone H4 (mono methyl K20) peptide (<a href='/en-us/products/unavailable/human-histone-h4-mono-methyl-k20-peptide-ab17043'>ab17043</a>)

Lane 7:

Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg with Histone H4 peptide (17-24) - di methyl K20

Secondary

All lanes:

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

Predicted band size: 11 kDa

Observed band size: 13 kDa

true

Exposure time: 90s

• WB

Project

Western blot - Human Histone H3 (tri methyl K4) peptide (AB1342)

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

Project1592****

Western blot - Human Histone H3 (tri methyl K4) peptide (AB1342)

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

Unknown

Western blot - Human Histone H3 (tri methyl K4) peptide (AB1342)

The peptide competition assay shows that this antibody is very specific for Histone H3 di-methylated at K27. It does not recognise Histone H3 mono, di, or tri methylated at K4, Histone H3 mono, di, or tri methylated at K9, or Histone H3 mono, or tri methylated at K27.

All lanes:

Western blot - Anti-Histone H3 (di methyl K27) antibody - ChIP Grade (<a href='/en-us/products/primary-antibodies/histone-h3-di-methyl-k27-antibody-chip-grade-ab24684'>ab24684</a>) at 1 µg/mL

Lanes 1, 3, 4, 5, 6, 7, 8, 9, 10 and 11:

Calf thymus histone lysate

Lane 2:

Calf thymus histone lysate with Human Histone H3 peptide (<a href='/en-us/products/unavailable/human-histone-h3-peptide-ab17163'>ab17163</a>)

Secondary

All lanes:

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

Predicted band size: 15 kDa

Observed band size: 17 kDa

false

Exposure time: 5s

• WB

Unknown

Western blot - Human Histone H3 (tri methyl K4) peptide (AB1342)

ab18521 recognises histone H3 as well as modified histone H3 at approximately 15 kDa(lane1). This is demonstrated by the blocking using the immunising unmodified histone H3 peptide (lane2), histone H3 mono methyl K4 (lane3), histone H3 di methyl K4 (lane4) and histone H3 tri methyl K4 (lane5).

All lanes:

Anti-Histone H3 antibody (ab31602) at 1 µg/mL

Lanes 1, 3, 4 and 5:

Calf thymus histone lysate at 20 µg

Lane 2:

Calf thymus histone lysate at 20 µg with Human Histone H3 peptide (<a href='/en-us/products/unavailable/human-histone-h3-peptide-ab14949'>ab14949</a>)

Predicted band size: 15 kDa

Observed band size: 15 kDa

false

• WB

Unknown

Western blot - Human Histone H3 (tri methyl K4) peptide (AB1342)

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

• WB

Project

Western blot - Human Histone H3 (tri methyl K4) peptide (AB1342)

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 (<a href='/en-us/products/primary-antibodies/histone-h4-tri-methyl-k20-antibody-chip-grade-ab9053'>ab9053</a>) at 1 µg/mL

Lanes 1, 5, 6 and 7:

Calf thymus histone lysate

Lane 2:

Calf thymus histone lysate with Human Histone H4 peptide (<a href='/en-us/products/unavailable/human-histone-h4-peptide-ab2622'>ab2622</a>)

Lane 3:

Calf thymus histone lysate with Human Histone H4 (mono methyl K20) peptide (<a href='/en-us/products/unavailable/human-histone-h4-mono-methyl-k20-peptide-ab17043'>ab17043</a>)

Lane 4:

Calf thymus histone lysate with Human Histone H4 (di methyl K20) peptide (<a href='/en-us/products/unavailable/human-histone-h4-di-methyl-k20-peptide-ab14964'>ab14964</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: 11 kDa

Observed band size: 13 kDa

false

• WB

Project

Western blot - Human Histone H3 (tri methyl K4) peptide (AB1342)

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 (<a href='/en-us/products/primary-antibodies/histone-h4-tri-methyl-k20-antibody-chip-grade-ab9053'>ab9053</a>) at 1 µg/mL

Lanes 1, 5, 6 and 7:

Calf thymus histone lysate

Lane 2:

Calf thymus histone lysate with Human Histone H4 peptide (<a href='/en-us/products/unavailable/human-histone-h4-peptide-ab2622'>ab2622</a>)

Lane 3:

Calf thymus histone lysate with Human Histone H4 (mono methyl K20) peptide (<a href='/en-us/products/unavailable/human-histone-h4-mono-methyl-k20-peptide-ab17043'>ab17043</a>)

Lane 4:

Calf thymus histone lysate with Human Histone H4 (di methyl K20) peptide (<a href='/en-us/products/unavailable/human-histone-h4-di-methyl-k20-peptide-ab14964'>ab14964</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: 11 kDa

Observed band size: 13 kDa

false

• WB

Unknown

Western blot - Human Histone H3 (tri methyl K4) peptide (AB1342)

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 (<a href='/en-us/products/primary-antibodies/histone-h3-mono-methyl-k4-antibody-chip-grade-ab8895'>ab8895</a>) at 1/500 dilution

All lanes:

Calf thymus histone lysate

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: 18 kDa

false

Exposure time: 2min

• WB

Lab

Western blot - Human Histone H3 (tri methyl K4) peptide (AB1342)

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

• Dot

AbReview21269****

Dot Blot - Human Histone H3 (tri methyl K4) peptide (AB1342)

ab1342 (at 0.01 and 0.1 μg) used in Dot Blot to test for cross-reactivity of ab8580 (1/2500). An HRP-conjugated Mouse anti-rabbit polyclonal (1/10000) was used as the secondary antibody.

This image is courtesy of an anonymous Abreview