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H3K27me3S28p | Histone H3 (trimethylated Lys27, p Ser28)

AS16 3184 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Chicken, C.elegans, D. melanogaster, Human, Mouse, plant, Rat, Xenopus sp.
H3K27me3S28p | Histone H3 (trimethylated Lys27, p Ser28) in the group Antibodies Plant/Algal  / DNA/RNA/Cell Cycle / Plant Epigenetics/DNA methylation at Agrisera AB (Antibodies for research) (AS16 3184)
H3K27me3S28p | Histone H3 (trimethylated Lys27, p Ser28)



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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 : 500-1 : 1000 (Dot), 1 : 200 (IF), 1 : 50 (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

ChIP using anti-H3K27me3S28p | Histone H3 (trimethylated Lys27, p Ser28)  polyclonal antibodies

Chromatin Immunoprecipitation: using anti-H3K27me3S28p antibodies. Chromatin from one million formaldehyde cross-linked HeLa cells was used with 2 μg of H3K27me3S28p and 20 μl of magnetic IgG beads per immunoprecipitation. A no antibody (No Ab) control was also used. Immunoprecipitated DNA was quantified using quantitative PCR and normalized to the input chromatin.

dot blot using anti-H3K27me3S28p | Histone H3 (trimethylated Lys27, p Ser28)  antibodies'

Dot Blot: using anti-H3K27me3S28p antibodies. Lane 1: S28p/K27 unmodified. Lane 2: S28p N-Term. Lane 3: S28p C-term. Lane 4: K27Me3. Lane 5: S28p/K27Me3. Load: 1, 10, and 100 picomoles of peptide. Primary antibody at 1 μg/ml 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-H3K27me3S28p | Histone H3 (trimethylated Lys27, p Ser28) polyclonal antibodies

Immunofluorescence: using anti-H3K27me3S28p antibodies. Tissue: HeLa cells. Fixation: 0.5% PFA. Primary antibody used at a 1:200 dilution for 1 h at RT. Secondary antibody: Dylight ®488 secondary antibody at 1:10 000 for 45 min at RT. Localization: Histone H3K27me3S28p is nuclear and chromosomal. Staining: Histone H3K27me3S28p is expressed in green, nuclei and alpha-tubulin are counterstained with DAPI (blue) and Dylight®550 (red).

western blot using anti-H3K27me3S28p | Histone H3 (trimethylated Lys27, p Ser28)  polyclonal antibodies

Western Blot: using anti-H3K27me3S28p antibodies. Lane 1: HeLa histone extracts. Lane 2: NIH-3T3 histone extracts. Lane 3: C. elegans embryo cell lysate. Load: 30 μg per lane. Primary antibody used at 1 μg/ml 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

Related products

Background

Background Common histone modifications include methylation of lysine and arginine, acetylation of lysine, phosphorylation of threonine and serine, and sumoylation, biotinylation, and ubiquitylation of lysine. Specifically, trimethylation of K27 is associated with gene silencing, whereas pS28 is associated with mitotis and immediate early genes.

Product citations

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 : 500-1 : 1000 (Dot), 1 : 200 (IF), 1 : 50 (IHC), 1 : 500 (WB)
calculated | apparent molecular mass [kDa]:

15 kDa

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
Picture (footer): application example

ChIP using anti-H3K27me3S28p | Histone H3 (trimethylated Lys27, p Ser28) polyclonal antibodies

Chromatin Immunoprecipitation: using anti-H3K27me3S28p antibodies. Chromatin from one million formaldehyde cross-linked HeLa cells was used with 2 μg of H3K27me3S28p and 20 μl of magnetic IgG beads per immunoprecipitation. A no antibody (No Ab) control was also used. Immunoprecipitated DNA was quantified using quantitative PCR and normalized to the input chromatin.

dot blot using anti-H3K27me3S28p | Histone H3 (trimethylated Lys27, p Ser28) antibodies'

Dot Blot: using anti-H3K27me3S28p antibodies. Lane 1: S28p/K27 unmodified. Lane 2: S28p N-Term. Lane 3: S28p C-term. Lane 4: K27Me3. Lane 5: S28p/K27Me3. Load: 1, 10, and 100 picomoles of peptide. Primary antibody at 1 μg/ml 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-H3K27me3S28p | Histone H3 (trimethylated Lys27, p Ser28) polyclonal antibodies

Immunofluorescence: using anti-H3K27me3S28p antibodies. Tissue: HeLa cells. Fixation: 0.5% PFA. Primary antibody used at a 1:200 dilution for 1 h at RT. Secondary antibody: Dylight ®488 secondary antibody at 1:10 000 for 45 min at RT. Localization: Histone H3K27me3S28p is nuclear and chromosomal. Staining: Histone H3K27me3S28p is expressed in green, nuclei and alpha-tubulin are counterstained with DAPI (blue) and Dylight®550 (red).

western blot using anti-H3K27me3S28p | Histone H3 (trimethylated Lys27, p Ser28) polyclonal antibodies

Western Blot: using anti-H3K27me3S28p antibodies. Lane 1: HeLa histone extracts. Lane 2: NIH-3T3 histone extracts. Lane 3: C. elegans embryo cell lysate. Load: 30 μg per lane. Primary antibody used at 1 μg/ml overnight at 4°C. Secondary antibody: IRDye800™ rabbit secondary antibody at 1:10 000 for 45 min at RT.
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: Common histone modifications include methylation of lysine and arginine, acetylation of lysine, phosphorylation of threonine and serine, and sumoylation, biotinylation, and ubiquitylation of lysine. Specifically, trimethylation of K27 is associated with gene silencing, whereas pS28 is associated with mitotis and immediate early genes.

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