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46439
PhosphoPlus® HDAC2 (Ser394) Antibody Duet
Primary Antibodies
Antibody Duet

PhosphoPlus® HDAC2 (Ser394) Antibody Duet #46439

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PhosphoPlus® HDAC2 (Ser394) Antibody Duet: Image 1
Flow cytometric analysis of Daudi cells, untreated (green) or treated with λ phosphatase (blue), using Phospho-HDAC2 (Ser394) (E8O2Z) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
PhosphoPlus® HDAC2 (Ser394) Antibody Duet: Image 2
CUT&RUN was performed with K-562 cells and either Phospho-HDAC2 (Ser394) (E8O2Z) Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using human NBPF1 promoter primers, human DTL exon 1 primers, and human ROBO1 promoter primers. 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.
PhosphoPlus® HDAC2 (Ser394) Antibody Duet: Image 3
CUT&RUN was performed with K-562 cells and either Phospho-HDAC2 (Ser394) (E8O2Z) Rabbit mAb or HDAC2 (D6S5P) Rabbit mAb #57156, using CUT&RUN Assay Kit #86652. DNA libraries were prepared using DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795. The figure shows binding across NBPF1 gene, a known target gene of phospho-HDAC2 (see additional figure containing CUT&RUN-qPCR data).
PhosphoPlus® HDAC2 (Ser394) Antibody Duet: Image 4
CUT&RUN was performed with K-562 cells and either Phospho-HDAC2 (Ser394) (E8O2Z) Rabbit mAb or HDAC2 (D6S5P) Rabbit mAb #57156, using CUT&RUN Assay Kit #86652. DNA libraries were prepared using DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 1 (upper), including NBPF1 (lower), a known target gene of phospho-HDAC2 (see additional figure containing CUT&RUN-qPCR data).
PhosphoPlus® HDAC2 (Ser394) Antibody Duet: Image 5
Western blot analysis of extracts from various cell lines using HDAC2 (D6S5P) Rabbit mAb.
PhosphoPlus® HDAC2 (Ser394) Antibody Duet: Image 6
Western blot analysis of extracts from various cell lines, untreated (-; left) or treated with calf intestinal phosphatase (CIP) (+; right), using Phospho-HDAC2 (Ser394) (E8O2Z) Rabbit mAb (upper) or HDAC2 (D6S5P) Rabbit mAb #57156 (lower).
PhosphoPlus® HDAC2 (Ser394) Antibody Duet: Image 7
Immunoprecipitation of HDAC2 from HeLa cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is HDAC2 (D6S5P) Rabbit mAb. Western blot analysis was performed using HDAC2 (D6S5P) Rabbit mAb.
PhosphoPlus® HDAC2 (Ser394) Antibody Duet: Image 8
Immunoprecipitation of phospho-HDAC2 protein from HeLa cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Phospho-HDAC2 (Ser394) (E8O2Z) Rabbit mAb. Western blot analysis was performed using Phospho-HDAC2 (Ser394) (E8O2Z) Rabbit mAb. Mouse Anti-Rabbit IgG (Light-Chain Specific) (D4W3E) mAb (HRP Conjugate) #93702 was used as a secondary antibody.
PhosphoPlus® HDAC2 (Ser394) Antibody Duet: Image 9
Confocal immunofluorescent analysis of HeLa cells using HDAC2 (D6S5P) Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red).
PhosphoPlus® HDAC2 (Ser394) Antibody Duet: Image 10
Chromatin immunoprecipitations were performed with cross-linked chromatin from K-562 cells and either HDAC2 (D6S5P) Rabbit mAb or HDAC1 (D5C6U) XP® Rabbit mAb #34589, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. HDAC2 and HDAC1 are known to have similar binding pattern on chromatin. The figure shows binding of both HDAC2 and HDAC1 across ACVR1 gene. For additional ChIP-seq tracks, please download the product data sheet.
PhosphoPlus® HDAC2 (Ser394) Antibody Duet: Image 11
Chromatin immunoprecipitations were performed with cross-linked chromatin from K-562 cells and either HDAC2 (D6S5P) Rabbit mAb or HDAC1 (D5C6U) XP® Rabbit mAb #34589, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. HDAC2 and HDAC1 are known to have similar binding pattern on chromatin. The figure shows binding of both HDAC2 and HDAC1 across chromosome 2 (upper), including ACVR1 gene (lower).
PhosphoPlus® HDAC2 (Ser394) Antibody Duet: Image 12
Chromatin immunoprecipitations were performed with cross-linked chromatin from K562 cells and either HDAC2 (D6S5P) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human RPL30 Exon 3 Primers #7014, human BCR exon 1 primers, and SimpleChIP® Human Nanog Promoter Primers #95064. 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.
To Purchase # 46439
Cat. # Size Qty. Price
46439S
1 Kit
$ 605

Product Includes Quantity Reactivity MW(kDa) Isotype
HDAC2 (D6S5P) Rabbit mAb 57156 100 µl H M R Mk 60 Rabbit IgG
Phospho-HDAC2 (Ser394) (E8O2Z) Rabbit mAb 69238 100 µl H M R Mk 62 Rabbit IgG

Product Description

PhosphoPlus® Duets from Cell Signaling Technology (CST) provide a means to assess protein activation status. Each Duet contains an activation-state and total protein antibody to your target of interest. These antibodies have been selected from CST's product offering based on superior performance in specified applications.
MW (kDa) 62

Background

Acetylation of the histone tail causes chromatin to adopt an "open" conformation, allowing increased accessibility of transcription factors to DNA. The identification of histone acetyltransferases (HATs) and their large multiprotein complexes has yielded important insights into how these enzymes regulate transcription (1,2). HAT complexes interact with sequence-specific activator proteins to target specific genes. In addition to histones, HATs can acetylate nonhistone proteins, suggesting multiple roles for these enzymes (3). In contrast, histone deacetylation promotes a "closed" chromatin conformation and typically leads to repression of gene activity (4). Mammalian histone deacetylases can be divided into three classes on the basis of their similarity to various yeast deacetylases (5). Class I proteins (HDACs 1, 2, 3, and 8) are related to the yeast Rpd3-like proteins, those in class II (HDACs 4, 5, 6, 7, 9, and 10) are related to yeast Hda1-like proteins, and class III proteins are related to the yeast protein Sir2. Inhibitors of HDAC activity are now being explored as potential therapeutic cancer agents (6,7).
Ser394 is phosphorylated by CK2, which promotes enzymatic activity and regulates complex formation (8).
  1. Marmorstein, R. (2001) Cell Mol Life Sci 58, 693-703.
  2. Gregory, P.D. et al. (2001) Exp Cell Res 265, 195-202.
  3. Liu, Y. et al. (2000) Mol Cell Biol 20, 5540-53.
  4. Cress, W.D. and Seto, E. (2000) J Cell Physiol 184, 1-16.
  5. Gray, S.G. and Ekström, T.J. (2001) Exp Cell Res 262, 75-83.
  6. Thiagalingam, S. et al. (2003) Ann. N.Y. Acad. Sci. 983, 84-100.
  7. Vigushin, D.M. and Coombes, R.C. (2004) Curr Cancer Drug Targets 4, 205-18.
  8. Tsai, S.C. and Seto, E. (2002) J Biol Chem 277, 31826-33.

Pathways & Proteins

Explore pathways + proteins related to this product.

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.

For Research Use Only. Not For Use In Diagnostic Procedures.
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