H3T3pK4me2 | Histone H3 (dimethylated Lys4, p Thr3)
AS16 3169 | Clonality: Polyclonal | Host: Rabbit | Reactivity: 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
Affinity purified serum
Format
Liquid
Quantity
50 µg
Storage
Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.
Tested applications
Dot blot (Dot), Chromatin immunoprecipitation (ChIP), Immunofluorescence (IF), Immunohistochemistry (IHC), Western blot (WB)
Recommended dilution
2-5 µg/milion cells (ChiP), 1 : 1000 (Dot), 1 : 50 (IF), 1 : 50 (IHC), 1 : 500 (WB)
Expected | apparent MW
15 kDa
Reactivity
Confirmed reactivity
Human
Predicted reactivity
Caenorhabditis elegans, 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 usingH3T3pK4me2 antibodies. Chromatin from one million formaldehyde cross-linked Hela cells was used with 2 μg of H3T3pK4me2 and 20 μl of magnetic beads per immunoprecipitation. A no antibody (No Ab) control was also used. Immunoprecipitated DNA was quantified using quantitative real-time PCR and normalized to the input chromatin.
Immunofluorescence using anti-H3T3pK4me2 antibodies. Tissue: HeLa cells. Fixation: 0.5% PFA. Primary antibody was used at a 1:50 dilution for 1 h at RT. Secondary antibody: Dylight® 488 secondary antibody at 1:10 000 for 45 min at RT. Localization: Histone H3T3pK4me2 is nuclear and chromosomal. Staining: H3T3pK4me2 is expressed in green while the nuclei and aplpha-tubulin were coexpressed with DAPI (blue) and Dylight® 550 (red).
Western Blot using anti-H3T3pK4me2 antibodies. 30 μg of NIH-3T3 histone extracts. 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 usingH3T3pK4me2 antibodies. Chromatin from one million formaldehyde cross-linked Hela cells was used with 2 μg of H3T3pK4me2 and 20 μl of magnetic beads per immunoprecipitation. A no antibody (No Ab) control was also used. Immunoprecipitated DNA was quantified using quantitative real-time PCR and normalized to the input chromatin.
Immunofluorescence using anti-H3T3pK4me2 antibodies. Tissue: HeLa cells. Fixation: 0.5% PFA. Primary antibody was used at a 1:50 dilution for 1 h at RT. Secondary antibody: Dylight® 488 secondary antibody at 1:10 000 for 45 min at RT. Localization: Histone H3T3pK4me2 is nuclear and chromosomal. Staining: H3T3pK4me2 is expressed in green while the nuclei and aplpha-tubulin were coexpressed with DAPI (blue) and Dylight® 550 (red).
Western Blot using anti-H3T3pK4me2 antibodies. 30 μg of NIH-3T3 histone extracts. 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
Methylation of lysine 4 on H3 (H3K4Me) and phosphorylation of threonine 3 (H3T3p) are known marks of transcriptional activation and mitosis. H3K4 has many known modifying enzymes (Set1, Set7/9, MLL, ASH1).
Product citations
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