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Anti-Histone H3 (mono methyl K18) HIST1H3A Rabbit Monoclonal Antibody

Figure 1. Western blot analysis of Histone H3 (mono methyl K18) using anti-Histone H3 (mono methyl K18) antibody (M12477-6).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human 293T whole cell lysates,
Lane 3: human CACO-2 whole cell lysates,
Lane 4: human HepG2 whole cell lysates,
Lane 5: human PC-12 whole cell lysates,
Lane 6: rat C6 whole cell lysates,
Lane 7: mouse RAW264.7 whole cell lysates,
Lane 8: mouse NIH/3T3 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Histone H3 (mono methyl K18) antigen affinity purified monoclonal antibody (Catalog # M12477-6) at 1:500 overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Histone H3 (mono methyl K18) at approximately 17 kDa. The expected band size for Histone H3 (mono methyl K18) is at 15 kDa.
Figure 2. IHC analysis of Histone H3 (mono methyl K18) using anti-Histone H3 (mono methyl K18) antibody (M12477-6).
Histone H3 (mono methyl K18) was detected in a paraffin-embedded section of mouse brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:500 rabbit anti-Histone H3 (mono methyl K18) Antibody (M12477-6) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Figure 3. IHC analysis of Histone H3 (mono methyl K18) using anti-Histone H3 (mono methyl K18) antibody (M12477-6).
Histone H3 (mono methyl K18) was detected in a paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:500 rabbit anti-Histone H3 (mono methyl K18) Antibody (M12477-6) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Figure 4. IF analysis of Histone H3 (mono methyl K18) using anti-Histone H3 (mono methyl K18) antibody (M12477-6) and anti-Beta Tubulin antibody (M01857-3).
Histone H3 (mono methyl K18) was detected in immunocytochemical section of U2OS cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated at 1:100 with rabbit anti-Histone H3 (mono methyl K18) Antibody (M12477-6) and mouse anti-Beta Tubulin antibody (M01857-3) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) and DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

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Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	WB, IHC, IF, ICC
Primary Accession	P68431
Host	Rabbit
Isotype	Rabbit IgG
Reactivity	Rat, Human, Mouse
Clonality	Monoclonal
Format	Liquid
Description	Anti-Histone H3 (mono methyl K18) HIST1H3A Rabbit Monoclonal Antibody . Tested in WB, IHC, ICC/IF applications. This antibody reacts with Human, Mouse, Rat.

Additional Information
Gene ID	8350;8351;8352;8353;8354;8355;8356;8357;8358;8968
Other Names	Histone H3.1, Histone H3/a, Histone H3/b, Histone H3/c, Histone H3/d, Histone H3/f, Histone H3/h, Histone H3/i, Histone H3/j, Histone H3/k, Histone H3/l, H3C1 (HGNC:4766), H3FA, HIST1H3A
Calculated MW	15404 Da
Application Details	WB 1:500-1:2000 IHC 1:500-1:2000 ICC/IF 1:100-1:500
Subcellular Localization	Nucleus. Chromosome.
Source	Eukaryota
Contents	Rabbit IgG in phosphate buffered saline, pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol, 0.4-0.5mg/ml BSA.
Clone Names	Clone: DFC-8
Immunogen	A synthesized peptide derived from human Histone H3 (mono methyl K18)
Purification	Affinity-chromatography
Storage	Store at -20°C for one year. For short term storage and frequent use, store at 4°C for up to one month. Avoid repeated freeze-thaw cycles.

For Research Use Only. Not For Use In Diagnostic Procedures.

Protein Information
Name	H3C1 (HGNC:4766)
Synonyms	H3FA, HIST1H3A
Function	Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling.
Cellular Location	Nucleus. Chromosome.

Research Areas

Application Protocols

Provided below are standard protocols that you may find useful for product applications.

Western Blot Protocol

Blocking Peptides Protocol

Dot Blot Protocol

Immunohistochemistry Protocol

Immunofluorescence Protocol

Immunoprecipitation Protocol

Flow Cytomety Protocol

Cell Culture Protocol