Revision 3

#41328Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IF-IC, ChIP

REACTIVITY:

H Mk

SENSITIVITY:

Endogenous

MW (kDa):

30

Source/Isotype:

Rabbit IgG

UniProt ID:

#P10412

Entrez-Gene Id:

3008

Product Information

Product Usage Information

For optimal ChIP results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

Application Dilution
Western Blotting 1:1000
Immunofluorescence (Immunocytochemistry) 1:200 - 1:800
Chromatin IP 1:50

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

Histone H1.4 (D4J5Q) Rabbit mAb recognizes endogenous levels of total histone H1.4 protein. This antibody also cross reacts with histone H1.5 (UniProt P16401) and weakly with histones H1.1 (UniProt Q02539), H1.2 (UniProt P16403), and H1.3 (UniProt P16402).

Species Reactivity:

Human, Monkey

Species predicted to react based on 100% sequence homology

Hamster, Bovine, Dog

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala176 of human histone H1.4 protein.

Background

Histones are evolutionarily conserved proteins that play a vital role in the compaction, storage, and regulation of DNA within the eukaryotic nucleus. The basic subunit of chromatin, the nucleosome core particle, is composed of DNA wound around two copies each of the core histone proteins H2A, H2B, H3, and H4 (1-3). Formation of higher order chromatin structure is facilitated through the binding of linker histone H1 to the nucleosome particle (chromatosome) (4-6). In humans and mice, there are 11 distinct histone H1 variants, which include the somatic variants (H1.1, H1.2, H1.3, H1.4, and H1.5) that are expressed ubiquitously, and cell type specific variants such as H1t found in the testis and H1.0 expressed in terminally differentiated cells (6-10). Binding of histone H1 to chromatin limits accessibility of DNA to other proteins by stabilizing nucleosome positioning, competing for binding sites, and limiting the activity of chromatin remodeling proteins such as the SWI/SNF complex (6,11-14). Histone H1 binding is highly dynamic and is thought to be regulated by post-translational modifications (6). For example, cell cycle regulated phosphorylation of histone H1 leads to chromatin condensation and decondensation depending on the site of phosphorylation and histone H1.4 Lys34 acetylation by GCN5 has been linked to increased mobility of H1.4 and transcriptional activation (6, 15-18).

  1. Kornberg, R.D. (1974) Science 184, 868-71.
  2. Kornberg, R.D. and Thomas, J.O. (1974) Science 184, 865-8.
  3. Workman, J.L. and Kingston, R.E. (1998) Annu Rev Biochem 67, 545-79.
  4. Thoma, F. et al. (1979) J Cell Biol 83, 403-27.
  5. Carruthers, L.M. et al. (1998) Biochemistry 37, 14776-87.
  6. Harshman, S.W. et al. (2013) Nucleic Acids Res 41, 9593-609.
  7. Eick, S. et al. (1989) Eur J Cell Biol 49, 110-5.
  8. Carozzi, N. et al. (1984) Science 224, 1115-7.
  9. Drabent, B. et al. (1991) Gene 103, 263-8.
  10. Pehrson, J.R. and Cole, R.D. (1982) Biochemistry 21, 456-60.
  11. Bustin, M. et al. (2005) Mol Cell 17, 617-20.
  12. Hill, D.A. and Imbalzano, A.N. (2000) Biochemistry 39, 11649-56.
  13. Ramachandran, A. et al. (2003) J Biol Chem 278, 48590-601.
  14. Horn, P.J. et al. (2002) Nat Struct Biol 9, 263-7.
  15. Hohmann, P. et al. (1976) J Biol Chem 251, 3685-92.
  16. D'Anna, J.A. et al. (1978) Nucleic Acids Res 5, 3195-207.
  17. Matsumoto, Y. et al. (1980) Nature 284, 181-3.
  18. Kamieniarz, K. et al. (2012) Genes Dev 26, 797-802.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IF-IC: Immunofluorescence (Immunocytochemistry) ChIP: Chromatin IP

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意，否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件，除非书 CST 的合法授书代表以书面形式书独接受， 否书均被拒书，并且无效。

专品专有“专供研究使用”的专专或专似的专专声明， 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的， 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专，且专用于研专用途。将专品用于专断、专防或治专目的， 或专专售（专独或作专专成）或其他商专目的而专专专品，均需要 CST 的专独专可。客专：(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品，或使用专品制造任何商专专品，(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专，或使用专品开专任何与 CST 的专品或服专专争的专品或服专， (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专，(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品， (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Orders: 877-616-CELL (2355)•[email protected]• Support: 877-678-TECH (8324)•[email protected]• Web: cellsignal.com
仅供研究使用。不得用于诊断流程。