Immunofluorescent analysis of Phospho-Histone H3 pSer10 (green) in HeLa cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed without (left panel) or with (right panel) a Phospho-Histone H3 polyclonal antibody (Product # PA5-17869) at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-rabbit IgG secondary antibody (Product # 35552) at a dilution of 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with DyLight 554 Phalloidin (Product # 21834) and nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan or a ToxInsight Instrument at 20X magnification.
|Tested species reactivity||Fruit fly, Human, Mouse, Non-human primate, Rat, Xenopus, Yeast|
|Published species reactivity||Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic phosphopeptide corresponding to residues surrounding pSer10 of human histone H3|
|Purification||Antigen affinity chromatography|
|Storage buffer||0.01M HEPES, pH 7.5, with 100µg/ml BSA, 0.15M NaCl, 50% glycerol|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:50|
|Immunohistochemistry (Frozen) (IHC (F))||1:400|
|Immunohistochemistry (Paraffin) (IHC (P))||1:200|
|Western Blot (WB)||1:1000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Immunocytochemistry (ICC)||See 1 publications below|
Antibodies to this protein (and modification) were previously sold as part of a Thermo Scientific Cellomics High Content Screening Kit. This replacement antibody is now recommended for researchers who need an antibody for high content cell based assays. It has been thoroughly tested and validated for cellular immunofluorescence (IF) applications. Further optimization including the selection of the most appropriate fluorescent Dylight conjugated secondary antibody may have to be performed for your high content assay.
It is not recommended to aliquot this antibody.
This antibody is not cross-reactive with other phosphorylated histones or with acetylated histones.
Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. This structure consists of approximately 146 bp of DNA wrapped around a nucleosome, an octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. This gene is intronless and encodes a member of the histone H3 family. Transcripts from this gene lack polyA tails; instead, they contain a palindromic termination element. This gene is found in the large histone gene cluster on chromosome 6p22-p21.3.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
Autophagy protects ovarian cancer-associated fibroblasts against oxidative stress.
PA5-17869 was used in immunocytochemistry and western blot to investigate protection of ovarian cancer-associated fibroblasts against oxidative stress by autophagy
|Wang Q,Xue L,Zhang X,Bu S,Zhu X,Lai D||Cell cycle (Georgetown, Tex.) (15:1376)||2016|