H3T3pK4ac | Histone H3 (ac Lys4, p Thr3)

AS16 3170 | Clonality: Polyclonal | Host: Rabbit | Reactivity: C.elegans, D. melanogaster, Human, Mouse, plant, Rat, Xenopus sp.

H3T3pK4ac | Histone H3 (ac Lys4, p Thr3)

Product Information

Immunogen KLH-conjugated synthetic acetylated/phosphorylated peptide surrounding Lysine 4 and Threonine 3 of human Histone H3

Host Rabbit

Clonality Polyclonal

Purity Immunogen 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 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), Immunohistochemistry (IHC), Western blot (WB)

Recommended dilution 2-5 µg/million cells (ChIP), 1 : 1000 (Dot), 1 : 100 (IF), 1 : 50 (IHC), 1 : 500 (WB)

Expected | apparent MW

15 kDa

Reactivity

Confirmed reactivity Caenorhabditis elegans, Human

Predicted reactivity 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

dot blot with anti-phospho-acetyl-Histone H3 antibodies

Dot Blot using anti-H3T3pK4ac antibodies. Lane 1: T3p. Lane 2: T3pK4me1. Lane 3: T3pK4me2. Lane 4: T3pK4ac. Lane 5: T3pR2me2s. Lane 6: K4 unmodified. Lane 7: K4me1. Lane 8: K4me2. Lane 9: K4me3. Lane 10: K4ac. Load: 1, 10, and 100 picomoles of peptide. Primary antibody used at 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-phospho-acetyl-Histone H3 antibodies

Immunofluorescence using anti-H3T3pK4ac antibodies. Tissue: HeLa cells. Fixation: 0.5% PFA. Primary antibody used at a 1:100 dilution for 1 h at RT. Secondary antibody: Dylight® 488 secondary antibody at 1:10 000 for 45 min at RT. Localization: Histone H3T3pK4ac is nuclear and chromosomal. Staining: H3T3pK4ac is expressed in green while the nuclei and aplpha-tubulin were coexpressed with DAPI (blue) and Dylight® 550 (red).

Western Blot using anti-phospho-acetyl-Histone H3 antibodies (H3T3pK4ac). Lane 1: HeLa histone extracts. Lane 2. NIH-3T3 histone extracts. Lane 3: C. elegans embryo lysate. Load: 30 μg per lane. Primary antibody used at 1:500 overnight at 4°C. Secondary antibody: IRDye800™ rabbit secondary antibody at 1:10 00 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 Chromatin is the arrangement of DNA and proteins in which chromosomes are formed. Correspondingly, chromatin is formed from nucleosomes, which are comprised of a set of four histone proteins (H2A, H2B, H3, H4) wrapped with DNA. Chromatin is a very dynamic structure in which numerous post-translational modifications work together to activate or repress the availability of DNA to be copied, transcribed, or repaired. These marks decide which DNA will be open and commonly active (euchromatin) or tightly wound to prevent access and activation (heterochromatin). Common histone modifications include methylation of lysine and arginine, acetylation of lysine, phosphorylation of threonine and serine, and sumoylation, biotinylation, and ubiquitylation of lysine. Phosphorylation of threonine 3 (H3T3p) is a known mitotic marker and modified by the Haspin/Thr3 enzyme, while acetylation of lysine 4 (H3K4ac) on histone 3 is associated with transcriptional activation by Esa1.

Alternative names: H3.3B, H3 histone, family 3A, H3.3AH3F3H3F3B, histone H3.3, MGC87782, MGC87783, H3pT3/K4ac .

Product citations

Picture (footer):	application example Dot Blot using anti-H3T3pK4ac antibodies. Lane 1: T3p. Lane 2: T3pK4me1. Lane 3: T3pK4me2. Lane 4: T3pK4ac. Lane 5: T3pR2me2s. Lane 6: K4 unmodified. Lane 7: K4me1. Lane 8: K4me2. Lane 9: K4me3. Lane 10: K4ac. Load: 1, 10, and 100 picomoles of peptide. Primary antibody used at 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-H3T3pK4ac antibodies. Tissue: HeLa cells. Fixation: 0.5% PFA. Primary antibody used at a 1:100 dilution for 1 h at RT. Secondary antibody: Dylight® 488 secondary antibody at 1:10 000 for 45 min at RT. Localization: Histone H3T3pK4ac is nuclear and chromosomal. Staining: H3T3pK4ac is expressed in green while the nuclei and aplpha-tubulin were coexpressed with DAPI (blue) and Dylight® 550 (red). Western Blot using anti-phospho-acetyl-Histone H3 antibodies (H3T3pK4ac). Lane 1: HeLa histone extracts. Lane 2. NIH-3T3 histone extracts. Lane 3: C. elegans embryo lysate. Load: 30 μg per lane. Primary antibody used at 1:500 overnight at 4°C. Secondary antibody: IRDye800™ rabbit secondary antibody at 1:10 00 for 45 min at RT.
calculated \| apparent molecular mass [kDa]:	15 kDa
Clonality:	Polyclonal
Format:	Liquid
Host:	Rabbit
immunogen:	KLH-conjugated synthetic acetylated/phosphorylated peptide surrounding Lysine 4 and Threonine 3 of human Histone H3
Purity:	Immunogen affinity purified serum.
Quantity:	50 µg
recommended dilution:	2-5 µg/million cells (ChIP), 1 : 1000 (Dot), 1 : 100 (IF), 1 : 50 (IHC), 1 : 500 (WB)
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), Immunohistochemistry (IHC), Western blot (WB)
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
Confirmed reactivity:	Caenorhabditis elegans, Human
predicted reactivity:	Chicken, Drosophila melanogaster, Mouse, Plant, Rat, Xenopus sp.
not reactive in:	No confirmed exceptions from predicted reactivity are currently known
background:	Chromatin is the arrangement of DNA and proteins in which chromosomes are formed. Correspondingly, chromatin is formed from nucleosomes, which are comprised of a set of four histone proteins (H2A, H2B, H3, H4) wrapped with DNA. Chromatin is a very dynamic structure in which numerous post-translational modifications work together to activate or repress the availability of DNA to be copied, transcribed, or repaired. These marks decide which DNA will be open and commonly active (euchromatin) or tightly wound to prevent access and activation (heterochromatin). Common histone modifications include methylation of lysine and arginine, acetylation of lysine, phosphorylation of threonine and serine, and sumoylation, biotinylation, and ubiquitylation of lysine. Phosphorylation of threonine 3 (H3T3p) is a known mitotic marker and modified by the Haspin/Thr3 enzyme, while acetylation of lysine 4 (H3K4ac) on histone 3 is associated with transcriptional activation by Esa1. Alternative names: H3.3B, H3 histone, family 3A, H3.3AH3F3H3F3B, histone H3.3, MGC87782, MGC87783, H3pT3/K4ac .

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