Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade

Name	Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade
Supplier	Abcam
Catalog	ab1220
Prices	$404.00
Sizes	100 µg
Host	Mouse
Clonality	Monoclonal
Isotype	IgG2a
Clone	mAbcam 1220
Applications	ICC/IF ICC/IF IP WB FC IHC-F ELISA IHC-P ChIP
Species Reactivities	Mouse, Rat, Chicken, Bovine, Human, Xenopus, Arabidopsis thaliana, C. elegans, Drosophila, S. pombe, Corn, Marmoset, Rice, Sheep, Yeast
Antigen	Synthetic peptide corresponding to Human Histone H3 aa 1-100 (di methyl K9) conjugated to Keyhole Limpet Haemocyanin (KLH) (Cysteine residue)
Description	Mouse Monoclonal
Gene	HIST3H3
Conjugate	Unconjugated
Supplier Page	Shop

Product images

Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220) at 1 µg/ml + HeLa (Human epitherlial carcinoma cell line) Whole Cell Lysate at 20 µgSecondaryGoat polyclonal to Mouse IgG-H&L- Pre-Adsorbed (HRP) at 1/10000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.

ab1220 staining human kidney sections by IHC-P using EXPOSE IHC detection kit (ab80436). Formalin fixed paraffin embedded tissue sections were pre-treated using heat mediated antigen retrieval (using a pressure cooker) with sodium citrate buffer (pH6) for 30 mins. The section was incubated with ab1220, 5µg/ml, for 1 hour at room temperature. DAB was used as the chromogen and the section was counterstained with haematoxylin and mounted with DPX.

All lanes : Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)Lane 1 : Calf thymus histone lysateLane 2 : Calf thymus histone lysate with Histone H3 peptide - unmodified at 1 µg/mlLane 3 : Calf thymus histone lysate with Human Histone H3 (mono methyl K9) peptide (ab1771) at 1 µg/mlLane 4 : Calf thymus histone lysate with Human Histone H3 (di methyl K9) peptide (ab1772) at 1 µg/mlLane 5 : Calf thymus histone lysate with Human Histone H3 (tri methyl K9) peptide (ab1773) at 1 µg/mlLane 6 : Calf thymus histone lysate with Human Histone H3 (di methyl K4) peptide (ab7768) at 1 µg/mlLane 7 : Calf thymus histone lysate with Human Histone H3 (di methyl K27) peptide (ab1781) at 1 µg/mlSecondaryRabbit Anti-Mouse IgG H&L (HRP) (ab6728) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.

Chromatin was prepared from U2OS cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The ChIP was performed with 25µg of chromatin, 2µg of ab1220 (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.

Interphase HeLa cells were fixed in paraformaldehyde and labelled with ab1220 (1/500). The staining is shown in green. The cells were counterstained with DAPI (red).

ELISA using ab1220 at varying antibody concentrations.The purple line indicates binding to the di methyl K9 peptide ab1772. Binding to the following peptides was not seen:unmodified K9 (ab2903),mono methyl K9 (ab1771),tri methyl K9 (ab1773),di methyl K27 (ab1781),tri methyl K27 peptide (ab1782),mono methyl K4 (ab1340),di methyl K4 (ab7768),tri methyl K4 (ab1342).This indicates the specificity of ab1220 for di methyl K9 of Histone H3.

ab1220 staining mouse embryonic stem cells by flow cytometry (gated on all living cells). The cells were trypsinized and stained with the antibody at 1ug/1.5 x 105 cells in a permeabilization buffer. A Cy3® conjugated goat anti-mouse antibody was used as the secondary.See Abreview

ab1220 staining rat liver tissue sections by IHC-P. Sections were formaldehyde fixed and subjected to heat mediated antigen retrieval in citrate buffer pH 6.0 prior to blocking with 5% serum for 30 minutes at 20°C. The primary antibody was diluted 1/400 and incubated with the sample for 45 minutes at 20°C. A HRP-conjugated goat anti-mouse antibody was used as the secondary.See Abreview

All lanes : Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220) at 2.5 µg/mlLane 1 : Fruit fly embryo tissue lysate - nuclear at 4.5 µgLane 2 : Fruit fly embryo tissue lysate - nuclear at 1.5 µgLane 3 : Fruit fly embryo tissue lysate - nuclear at 0.5 µgSecondaryHRP-conjugated Goat anti-mouse IgG polyclonal at 1/2000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.

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