Revision 3

#69238

Store at -20C

CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, IP, FC-FP, C&R, C&T

Reactivity:
H M R Mk

Sensitivity:
Endogenous

MW (kDa):
62

Source/Isotype:
Rabbit IgG

UniProt ID:
#Q92769

Entrez-Gene Id:
3066

Product Usage Information

The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.

The CUT&Tag dilution was determined using CUT&Tag Assay Kit #77552.

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Flow Cytometry (Fixed/Permeabilized) 1:200 - 1:800
CUT&RUN 1:50
CUT&Tag 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

Phospho-HDAC2 (Ser394) (E8O2Z) Rabbit Monoclonal Antibody recognizes endogenous levels of HDAC2 protein only when phosphorylated at Ser394.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phophopeptide corresponding to residues surrounding Ser394 of human HDAC2 protein.

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).

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 IP: Immunoprecipitation FC-FP: Flow Cytometry (Fixed/Permeabilized) C&R: CUT&RUN C&T: CUT&Tag

Cross-Reactivity Key

H: Human M: Mouse R: Rat Mk: Monkey

Trademarks and Patents

Cell Signaling Technology is a 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.

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 3

Cell Signaling Technology Logo
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).
Western Blotting Image 1: Phospho-HDAC2 (Ser394) (E8O2Z) Rabbit Monoclonal Antibody
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.
Immunoprecipitation Image 1: Phospho-HDAC2 (Ser394) (E8O2Z) Rabbit Monoclonal Antibody
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.
Flow Cytometry Image 1: Phospho-HDAC2 (Ser394) (E8O2Z) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 3

Cell Signaling Technology Logo
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).
CUT & RUN Image 1: Phospho-HDAC2 (Ser394) (E8O2Z) Rabbit Monoclonal Antibody
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).
CUT & RUN Image 2: Phospho-HDAC2 (Ser394) (E8O2Z) Rabbit Monoclonal Antibody
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.
CUT & RUN Image 3: Phospho-HDAC2 (Ser394) (E8O2Z) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 3

Cell Signaling Technology Logo
CUT&Tag was performed with K-562 cells and Phospho-HDAC2 (Ser394) (E8O2Z) Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figure shows binding across the NBPF1 gene.
CUT & Tag Image 1: Phospho-HDAC2 (Ser394) (E8O2Z) Rabbit Monoclonal Antibody
CUT&Tag was performed with K-562 cells and Phospho-HDAC2 (Ser394) (E8O2Z) Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figures show binding across chromosome 1 (upper), including the NBPF1 gene (lower).
CUT & Tag Image 2: Phospho-HDAC2 (Ser394) (E8O2Z) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.