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Anti-Methyl-Histone H3 (di K4) HIST1H3A Rabbit Monoclonal Antibody

Immunohistochemical analysis of paraffin-embedded human liver, using Methyl-Histone H3 (di K4) Antibody(M12477-1)
HIST1H3A was detected in paraffin-embedded tissue section. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-HIST1H3A Antibody (M12477-1)overnight at 4℃. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37℃. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Figure 1. Western blot analysis of Methyl-Histone H3 (di K4) using anti-Methyl-Histone H3 (di K4) antibody (M12477-1).
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: rat 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-Methyl-Histone H3 (di K4) antigen affinity purified monoclonal antibody (Catalog # M12477-1) 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 1:500 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 Methyl-Histone H3 (di K4) at approximately 15 kDa. The expected band size for Methyl-Histone H3 (di K4) is at 15 kDa.

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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, IP, FC
Primary Accession	P68431
Host	Rabbit
Isotype	Rabbit IgG
Reactivity	Rat, Human, Mouse
Clonality	Monoclonal
Format	Liquid
Description	Anti-Methyl-Histone H3 (di K4) HIST1H3A Rabbit Monoclonal Antibody . Tested in WB, IHC, ICC/IF, IP, Flow Cytometry 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 1:50-1:200 ICC/IF 1:50-1:200 IP 1:50 FC 1:50
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: AFF-8
Immunogen	A synthesized peptide derived from human Methyl-Histone H3 (di K4)
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