Western blot analysis was performed on acid extracts (30µg lysate) of HEK-293 (Lane 1), HEL 92.1.7 (Lane 2), K562 (Lane 3), PC-12 (Lane 4) and THP-1 (Lane 5). The blot was probed with Anti-Histone H3 Rabbit Polyclonal Antibody (Product # PA1-16941, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25µg/ml, 1:4000 dilution). A 17 kDa band corresponding to Histone H3 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide between amino acids 100-135 of the human Histone H3 protein.|
|Purification||Ammonium sulfate precipitation|
|Contains||0.02% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:1,000|
* 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.
This antibody is expected to cross react with all mammals and a wide range of other species based on sequence homology.
Suggested positive control: Hela whole cell lysate, MEF and HeLa lysates, MEF whole cell lysate0.
In diploid eukaryotic cells, the chromatin fibers are about 20nM in diameter. They consist of two major components in equal amounts, DNA and basic proteins called histones. Histones are believed to be regularly arranged in the deep grove of the DNA helix. The recurring positive charges of the histones form electrostatic associations with the negatively charged phosphate groups of DNA making the DNA more stable and flexible. This allows for the supercoiling of the chromatin fibers.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.