H3K18me2 | Histone H3 (dimethylated Lys18)
AS16 3182 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Chicken, C.elegans, D. melanogaster, Human, Mouse, plant, Rat, Xenopus sp.
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Product Information
Immunogen
KLH-conjugated synthetic peptide
Host
Rabbit
Clonality
Polyclonal
Purity
Immunogen affinity purified serum.
Format
Liquid
Quantity
50 ĩg
Storage
Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
Tested applications
Chromatin immunoprecipitation (ChIP), Dot blot (Dot), Immunofluorescence (IF), Western blot (WB)
Recommended dilution
2-5 µg/million cells (ChIP), 1 : 1000 (Dot), 1 : 500 (IF), 1 : 100 (IHC), 1 : 500 (WB)
Expected | apparent MW
15 kDa
Reactivity
Confirmed reactivity
Human
Predicted reactivity
Caenorhabditis elegans, Chicken, Drosophila melanogaster, Mouse, Plant, Rat, Xenopus sp.
Not reactive in
No confirmed exceptions from predicted reactivity are currently known
Application examples
Application examples
application example
Chromatin Immunoprecipitation using anti-H3K18me2 antibodies. Chromatin from one million formaldehyde cross-linked Hela cells was used with 2 μg of H3K18me2 antibody and 20 μl of magnetic IgG beads per immunoprecipitation. A no antibody (No Ab) control was also used. Immunoprecipitated DNA was quantified using real-time PCR and normalized to the input chromatin.
Dot Blot using anti-H3K18me2 antibodies. Lane 1: K18 unmodified. Lane 2: K18Me2. Lane 3: K18Me3. Lane 4: K18ac. Lane 5: K18Me1. Load: 0.1, 0.3, and 1 picomoles of peptide. Primary antibody used at a 1:1 000 dilution for 45 min at 4°C. Secondary antibody: Dylight®488 rabbit secondary antibody at 1:10 000 for 45 min at RT.
Immunofluorescence using anti-H3K18me2 antibodies. Tissue: HeLa cells. Fixation: 0.5% PFA. Primary antibody used at a 1:500 dilution for 1 h at RT. Secondary antibody: Dylight® 488 secondary antibody at 1:10 000 for 45 min at RT. Localization: Histone H3K18me2 is nuclear and chromosomal. Staining: Histone H3K18me2 is expressed in green, nuclei and alpha-tubulin are counterstained with DAPI (blue) and Dylight® 594 (red).
Western Blot using anti-H3K18me2 antibodies. 30 μg of NIH-3T3 histone extract. Primary antibody used at a 1:500 dilution overnight at 4°C. Secondary antibody: IRDye800™ rabbit secondary antibody at 1:10 000 for 45 min at RT.
Chromatin Immunoprecipitation using anti-H3K18me2 antibodies. Chromatin from one million formaldehyde cross-linked Hela cells was used with 2 μg of H3K18me2 antibody and 20 μl of magnetic IgG beads per immunoprecipitation. A no antibody (No Ab) control was also used. Immunoprecipitated DNA was quantified using real-time PCR and normalized to the input chromatin.
Dot Blot using anti-H3K18me2 antibodies. Lane 1: K18 unmodified. Lane 2: K18Me2. Lane 3: K18Me3. Lane 4: K18ac. Lane 5: K18Me1. Load: 0.1, 0.3, and 1 picomoles of peptide. Primary antibody used at a 1:1 000 dilution for 45 min at 4°C. Secondary antibody: Dylight®488 rabbit secondary antibody at 1:10 000 for 45 min at RT.
Immunofluorescence using anti-H3K18me2 antibodies. Tissue: HeLa cells. Fixation: 0.5% PFA. Primary antibody used at a 1:500 dilution for 1 h at RT. Secondary antibody: Dylight® 488 secondary antibody at 1:10 000 for 45 min at RT. Localization: Histone H3K18me2 is nuclear and chromosomal. Staining: Histone H3K18me2 is expressed in green, nuclei and alpha-tubulin are counterstained with DAPI (blue) and Dylight® 594 (red).
Western Blot using anti-H3K18me2 antibodies. 30 μg of NIH-3T3 histone extract. Primary antibody used at a 1:500 dilution overnight at 4°C. Secondary antibody: IRDye800™ rabbit secondary antibody at 1:10 000 for 45 min at RT.
Additional information
Additional information
This antibody preparation is provided in 20 mM Potassium Phosphate pH 7,2, 150 mM NaCl, 0,01% sodium azide and 30% glycerol
Background
Background
The di-methylated K18 on histone H3 is a seemingly transient post-translational modification. H3K18 is better known to be acetylated, and occasionally mono-methylated. Suv39h1, a well-studied histone methyltransferase seems to be responsible for the transition of acetylation and methylation at this H3 modification site. The di-methylated K18 on H3 seems to be associated with embryological development and possibly implantation.
Product citations
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