Immunofluorescence analysis of Histone H3 was performed using 70% confluent log phase NIH/3T3 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Histone H3 Rabbit Polyclonal Antibody (PA516183) at 1:250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Bovine, Dog, Chicken, Fruit fly, Horse, Human, Mouse, Non-human primate, Rat, Xenopus, Zebrafish|
|Published species reactivity||Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to the carboxy-terminal sequence of human histone H3|
|Purification||Antigen affinity chromatography|
|Storage buffer||0.01M HEPES, pH 7.5, with 0.15M NaCl, 100µg/ml BSA, 50% glycerol|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||1.5 µg/10^6 cells|
|Flow Cytometry (Flow)||1:20|
|Western Blot (WB)||1:1,000 - 1:4,000|
* 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.
|Western Blot (WB)||See 1 publications below|
It is not recommended to aliquot this antibody.
This antibody is not cross-reactive with other histones.
Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Two molecules of each of the four core histones (H2A, H2B, H3, and H4) form an octamer, around which approximately 146 bp of DNA is wrapped in repeating units, called nucleosomes. The linker histone, H1, interacts with linker DNA between nucleosomes and functions in the compaction of chromatin into higher order structures. This gene is intronless and encodes a member of the histone H3 family. Transcripts from this gene lack polyA tails but instead contain a palindromic termination element. This gene is found in the large histone gene cluster on chromosome 6.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
A new class of cyclin dependent kinase in Chlamydomonas is required for coupling cell size to cell division.
PA5-16183 was used in western blot to determine the requirement of coupling cell size to cell division by a new class of cyclin dependent kinase in chlamydomonas
|Li Y,Liu D,López-Paz C,Olson BJ,Umen JG||eLife (5:null)||2016|