Antibodies, Proteins, Kits and Reagents for Life Science

Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab11946)

Key features and details

Goat polyclonal to Histone H3 (di methyl K4) - ChIP Grade
Suitable for: IHC-P, ChIP, WB
Reacts with: Mouse, Rat, Cow, Human
Isotype: IgG

Product name

Anti-Histone H3 (di methyl K4) antibody - ChIP Grade
See all Histone H3 primary antibodies
Description

Goat polyclonal to Histone H3 (di methyl K4) - ChIP Grade
Host species

Goat

Tested Applications & Species

Application	Species
ChIP	Human
IHC-P	Human
WB	Mouse Rat Cow

See all applications and species data

Immunogen

Synthetic peptide within Human Histone H3 aa 1-100 (di methyl K4) conjugated to keyhole limpet haemocyanin. The exact sequence is proprietary.
(Peptide available as ab7768)
General notes

The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

Learn more here.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 1% BSA

This product may contain up to 3% BSA depending on the batch. For specific batch formulations please contact us.
Concentration information loading...
Purity

Immunogen affinity purified
Clonality

Polyclonal
Isotype

IgG
Research areas

ChIP - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab11946)

Chromatin was prepared from Hela 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 ab11946 (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). Primers and probes are located in the first kb of the transcribed region.
Western blot - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab11946)

All lanes : Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab11946) at 1 µg/ml

Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Rabbit polyclonal to Goat IgG - H&L - Pre-Adsorbed (HRP) (ab65486) at 1/3000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 15 kDa
Observed band size: 17 kDa
why is the actual band size different from the predicted?

Exposure time: 2 minutes
Western blot - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab11946)

Lane 1 : Marker
Lanes 2-12 : Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab11946) at 1 µg/ml

Lane 2 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) at 0.5 µg
Lane 3 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) at 0.5 µg with Human Histone H3 (unmodified) peptide (ab7228) at 0.75 µg/ml
Lane 4 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) at 0.5 µg with Human Histone H3 (mono methyl K4) peptide (ab1340) at 0.75 µg/ml
Lane 5 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) at 0.5 µg with Human Histone H3 (di methyl K4) peptide (ab7768) at 0.75 µg/ml
Lane 6 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) at 0.5 µg with Human Histone H3 (tri methyl K4) peptide (ab1342) at 0.75 µg/ml
Lane 7 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) at 0.5 µg with Human Histone H3 (mono methyl K9) peptide (ab1771) at 0.75 µg/ml
Lane 8 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) at 0.5 µg with Human Histone H3 (di methyl K9) peptide (ab1772) at 0.75 µg/ml
Lane 9 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) at 0.5 µg with Human Histone H3 (tri methyl K9) peptide (ab1773) at 0.75 µg/ml
Lane 10 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) at 0.5 µg with Human Histone H3 (mono methyl K27) peptide (ab1780) at 0.75 µg/ml
Lane 11 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) at 0.5 µg with Human Histone H3 (di methyl K27) peptide (ab1781) at 0.75 µg/ml
Lane 12 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) at 0.5 µg with Human Histone H3 (tri methyl K27) peptide (ab1782) at 0.75 µg/ml

Secondary
Lanes 2-12 : Rabbit polyclonal to Goat IgG (Alexa Fluor® 680) at 1/10000 dilution

Performed under reducing conditions.

Predicted band size: 15 kDa
Observed band size: 17 kDa why is the actual band size different from the predicted?
Western blot - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab11946)

Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab11946) at 1 µg/ml + PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

Performed under reducing conditions.

Predicted band size: 15 kDa
Observed band size: 17 kDa why is the actual band size different from the predicted?

Exposure time: 20 minutes
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab11946)

IHC image of Histone H3 staining in human normal cervix formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab11946, 1µg/ml, for 15 mins at room temperature. A donkey anti-goat biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

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