Analysis Note

ControlHeLa acid extracts

Application

Research Sub CategoryHistones

Immunoblot Analysis:
A 1:500-1:2000 dilution of this lot detected phosphorylated histone H3 in acid extracted proteins from mitotic HeLa cells treated with colcemid, but not unmodified recombinant histone H3 (Catalog # 14-494).

Western Blot Analysis:
A 1:5,000 dilution of a previous lot detected phosphorylated histone H3 in acid extracted proteins from mitotic HeLa cells treated with colcemid, but not unmodified recombinant histone H3 (Catalog # 14-494).


Multiplexing:
Beadlyte™ Histone-Peptide Specificity Assay: 1:1,000-1:50,000 dilutions of a previous lot were incubated with histone H3 peptides containing various modifications conjugated to Luminex™ microspheres. Only the peptide containing phospho-threonine 11 was
detected (Figure C).

Peptide Inhibition Analysis: 1 µM of histone H3 peptide containing phospho-threonine 11 abolished detection of histone H3 by anti-phospho-Histone H3 (Thr11), clone MC83 in immunoblot analysis of acid extracts from
colcemid treated HeLa cells (Figure D, lane 4).

Research CategoryEpigenetics & Nuclear Function

Anti-phospho-Histone H3 (Thr11) Antibody, clone MC83 is a rabbit monoclonal antibody for detection of phospho-Histone H3 (Thr11) also known as H3T11P, Histone H3 (phospho T11) & has been validated in WB, PIA, Mplex.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

Histones are highly conserved proteins that serve as the structural scaffold for the organization of nuclear DNA into chromatin. The four core histones, H2A, H2B, H3, and H4, assemble into an octamer (2 molecules of each). Histones are modified post-translationally by the actions of enzymes in both the nucleus and cytoplasm. These modifications, which include acetylation, phosphorylation, methylation, and ubiquitin-ation, occur predominantly on the N-terminal and C-terminal tails that extend beyond the nucleosome core particle. Phosphorylation at Ser10, Ser28 and Thr11 of histone H3 is tightly correlated with chromosome condensation during both mitosis and meiosis. In mammals, phosphorylation of Thr11 modification is restricted to the centromeric region while in plant cells, phosphorylation of Thr11 was distributed along the entire length of condensed chromosomes. Mitosis-specific phosphorylation of histone H3 at Thr11 is mediated by Dlk/ZIP kinase.

Immunogen

Peptide containing ...KS[pT]G..., in which pT corresponds to phospho-threonine at position 11 of human histone H3

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Linkage

Replaces: MABE468

Physical form

Cultured supernantant in 0.05% sodium azide

Quality

Routinely evaluated by immunoblot analysis.

Specificity

Broad species cross-reactivity is expected based on sequence homology. An additional unidentified band at ~40 kDa appears in some lysates.

Recognizes histone H3 phosphorylated at Thr11, Mr 17kDa.

Storage and Stability

Stable for 1 year at -20°C from date of receipt.
For maximum recovery of product, centrifuge the vial prior to removing the cap.

Target description

17kDa