Revision 5
#50722
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For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:
W, ChIP, ChIP-seq
Reactivity:
H M R Mk
Sensitivity:
Endogenous
MW (kDa):
14
Source/Isotype:
Rabbit IgG
UniProt ID:
#P0C0S5
Entrez-Gene Id:
3015
Product Usage Information
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
Chromatin IP | 1:50 |
Chromatin IP-seq | 1:50 |
Storage
Specificity/Sensitivity
H2A.Z (E9M5G) Rabbit mAb recognizes endogenous levels of total H2A.Z protein. This antibody does not cross-react with H2A protein.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Val110 of human H2A.Z protein.
Background
Modulation of chromatin structure plays a critical role in the regulation of transcription in eukaryotes. The nucleosome, made up of four core histone proteins (H2A, H2B, H3 and H4), is the primary building block of chromatin. In addition to the growing number of post-translational histone modifications regulating chromatin structure, cells can also exchange canonical histones with variant histones that can directly or indirectly modulate chromatin structure (1). There are five major variants of histone H2A: canonical H2A (most abundant), H2A.X, MacroH2A, H2ABbd and H2A.Z (2). Histone H2A.Z, the most conserved variant across species, functions as both a positive and negative regulator of transcription and is important for chromosome stability (2). Several homologous protein complexes, such as SWR-C (S. cerevisiae), TIP60 (D. melanogaster) and SRCAP (mammals), have been shown to catalyze the ATP-dependent exchange of H2A.Z for H2A in the nucleosome (3,4,5). This exchange of histone H2A variants changes histone-histone interactions in the nucleosome core and alters an acidic patch on the surface of the nucleosome, resulting in changes in nucleosome stability and binding of non-histone proteins such as HP1α (6,7).
Background References
- Jin, J. et al. (2005) Trends Biochem Sci 30, 680-7.
- Raisner, R.M. and Madhani, H.D. (2006) Curr Opin Genet Dev 16, 119-24.
- Mizuguchi, G. et al. (2004) Science 303, 343-8.
- Kusch, T. et al. (2004) Science 306, 2084-7.
- Ruhl, D.D. et al. (2006) Biochemistry 45, 5671-7.
- Suto, R.K. et al. (2000) Nat Struct Biol 7, 1121-4.
- Fan, J.Y. et al. (2004) Mol Cell 16, 655-61.
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
W: Western Blotting 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 G: Goat 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.
Limited Uses
Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.
Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.
Revision 5
#50722
H2A.Z (E9M5G) Rabbit mAb
Western blot analysis of HeLa, NIH/3T3, C6, and COS-7 cell lysates, in addition to 10 ng of recombinant H2A (rH2A) and H2A.Z (rH2A.Z) proteins, using Histone H2A.Z (E9M5G) Rabbit mAb (upper) and Histone H2A (D6O3A) Rabbit mAb #12349 (lower).
Chromatin immunoprecipitations were performed with cross-linked chromatin from A549 cells and H2A.Z (E9M5G) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA libraries were prepared using DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795. The figures show binding across MRPS5 gene.
Chromatin immunoprecipitations were performed with cross-linked chromatin from A549 cells and H2A.Z (E9M5G) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 22 (upper), including MRPS5 gene (lower).
Revision 5
#50722
H2A.Z (E9M5G) Rabbit mAb
Chromatin immunoprecipitations were performed with cross-linked chromatin from A549 cells and either H2A.Z (E9M5G) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using human SLC2A8 promoter primers, SimpleChIP® Human VAMP1 Promoter Primers #25921, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.