Revision 10

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877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

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

Applications:
W, IP, IHC-P, IF-IC, ChIP, ChIP-seq, C&R, C&T

Reactivity:
H M R Mk

Sensitivity:
Endogenous

MW (kDa):
62

Source/Isotype:
Rabbit IgG

UniProt ID:
#Q13547

Entrez-Gene Id:
3065

Product Usage Information

For optimal ChIP and ChIP-seq 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.

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
Immunohistochemistry (Paraffin) 1:50 - 1:200
Immunofluorescence (Immunocytochemistry) 1:50 - 1:200
Chromatin IP 1:50
Chromatin IP-seq 1:50
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.

For a carrier free (BSA and azide free) version of this product see product #51225.

Specificity/Sensitivity

HDAC1 (D5C6U) XP® Rabbit mAb recognizes endogenous levels of total HDAC1 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala440 of human HDAC1 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).

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 IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry) ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq 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.

SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.

XP 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.

Orders: 877-616-CELL (2355) orders@cellsignal.com Support: 877-678-TECH (8324) info@cellsignal.com Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 10

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using HDAC1 (D5C6U) XP® Rabbit mAb.
Western Blotting Image 1: HDAC1 (D5C6U) XP<sup>®</sup> Rabbit mAb
Immunoprecipitation of HDAC1 from HeLa cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is HDAC1 (D5C6U) XP® Rabbit mAb. Western blot analysis was performed using HDAC1 (D5C6U) Rabbit mAb.
Immunoprecipitation Image 1: HDAC1 (D5C6U) XP<sup>®</sup> Rabbit mAb

Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using HDAC1 (D5C6U) XP® Rabbit mAb.

Immunohistochemistry Image 1: HDAC1 (D5C6U) XP<sup>®</sup> Rabbit mAb
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For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 10

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Immunohistochemical analysis of paraffin-embedded human adenoid cystic carcinoma of the salivary gland using HDAC1 (D5C6U) XP® Rabbit mAb.

Immunohistochemistry Image 2: HDAC1 (D5C6U) XP<sup>®</sup> Rabbit mAb
Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using HDAC1 (D5C6U) XP® Rabbit mAb.
Immunohistochemistry Image 3: HDAC1 (D5C6U) XP<sup>®</sup> Rabbit mAb

Immunohistochemical analysis of paraffin-embedded human glassy cell carcinoma of the cervix using HDAC1 (D5C6U) XP® Rabbit mAb.

Immunohistochemistry Image 4: HDAC1 (D5C6U) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) orders@cellsignal.com Support: 877-678-TECH (8324) info@cellsignal.com Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 10

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human urothelial carcinoma using HDAC1 (D5C6U) XP® Rabbit mAb.
Immunohistochemistry Image 5: HDAC1 (D5C6U) XP<sup>®</sup> Rabbit mAb

Immunohistochemical analysis of paraffin-embedded normal human lymph node using HDAC1 (D5C6U) XP® Rabbit mAb.

Immunohistochemistry Image 6: HDAC1 (D5C6U) XP<sup>®</sup> Rabbit mAb

Immunohistochemical analysis of paraffin-embedded normal human testis using HDAC1 (D5C6U) XP® Rabbit mAb.

Immunohistochemistry Image 7: HDAC1 (D5C6U) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) orders@cellsignal.com Support: 877-678-TECH (8324) info@cellsignal.com Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 10

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded normal human small intestine (left) or normal human uterus (right) using HDAC1 (D5C6U) XP® Rabbit mAb (top) or an HDAC1 Mouse mAb (bottom). These two antibodies detect unique, non-overlapping epitopes on human HDAC1. The similar staining patterns obtained with both antibodies help to confirm the specificity of the staining.
Immunohistochemistry Image 8: HDAC1 (D5C6U) XP<sup>®</sup> Rabbit mAb
Immunohistochemical analysis of paraffin-embedded CT26.WT syngeneic tumor using HDAC1 (D5C6U) XP® Rabbit mAb.
Immunohistochemistry Image 9: HDAC1 (D5C6U) XP<sup>®</sup> Rabbit mAb
Immunohistochemical analysis of paraffin-embedded Renca syngeneic tumor using HDAC1 (D5C6U) XP® Rabbit mAb.
Immunohistochemistry Image 10: HDAC1 (D5C6U) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) orders@cellsignal.com Support: 877-678-TECH (8324) info@cellsignal.com Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 10

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded mouse brain using HDAC1 (D5C6U) XP® Rabbit mAb.
Immunohistochemistry Image 11: HDAC1 (D5C6U) XP<sup>®</sup> Rabbit mAb

Immunohistochemical analysis of paraffin-embedded mouse colon using HDAC1 (D5C6U) XP® Rabbit mAb.

Immunohistochemistry Image 12: HDAC1 (D5C6U) XP<sup>®</sup> Rabbit mAb

Immunohistochemical analysis of paraffin-embedded mouse liver using HDAC1 (D5C6U) XP® Rabbit mAb.

Immunohistochemistry Image 13: HDAC1 (D5C6U) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) orders@cellsignal.com Support: 877-678-TECH (8324) info@cellsignal.com Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 10

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded endometrioid adenocarcinoma using HDAC1 (D5C6U) XP® Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right). 
Immunohistochemistry Image 14: HDAC1 (D5C6U) XP<sup>®</sup> Rabbit mAb
Confocal immunofluorescent analysis of HeLa cells using HDAC1 (D5C6U) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red).
Immunofluorescence Image 1: HDAC1 (D5C6U) XP<sup>®</sup> Rabbit mAb
Chromatin immunoprecipitations were performed with cross-linked chromatin from K-562 cells and either HDAC2 (D6S5P) Rabbit mAb #57156 or HDAC1 (D5C6U) XP® Rabbit mAb, 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.
Chromatin Immunoprecipitation Image 1: HDAC1 (D5C6U) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) orders@cellsignal.com Support: 877-678-TECH (8324) info@cellsignal.com Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 10

Cell Signaling Technology Logo
Chromatin immunoprecipitations were performed with cross-linked chromatin from K-562 cells and either HDAC2 (D6S5P) Rabbit mAb #57156 or HDAC1 (D5C6U) XP® Rabbit mAb, 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).
Chromatin Immunoprecipitation Image 2: HDAC1 (D5C6U) XP<sup>®</sup> Rabbit mAb
Chromatin immunoprecipitations were performed with cross-linked chromatin from K-562 cells and either HDAC1 (D5C6U) XP® 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, SimpleChIP® Human GAPDH Exon 1 Primers #5516, 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.
Chromatin Immunoprecipitation Image 3: HDAC1 (D5C6U) XP<sup>®</sup> Rabbit mAb
CUT&RUN was performed with K562 cells and either HDAC1 (D5C6U) XP® Rabbit mAb or G9a/EHMT2 (D5R4R) XP® Rabbit mAb #68851, using CUT&RUN Assay Kit #86652. DNA libraries were prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figure shows binding across CTPS1, a known target gene of HDAC1 and G9a (see additional figure containing CUT&RUN-qPCR data).
CUT & RUN Image 1: HDAC1 (D5C6U) XP<sup>®</sup> Rabbit mAb
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For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 10

Cell Signaling Technology Logo
CUT&RUN was performed with K562 cells and either HDAC1 (D5C6U) XP® Rabbit mAb or G9a/EHMT2 (D5R4R) XP® Rabbit mAb #68851, using CUT&RUN Assay Kit #86652. DNA libraries were prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 1 (upper), including CTPS1 (lower), a known target gene of HDAC1 and G9a (see additional figure containing CUT&RUN-qPCR data).
CUT & RUN Image 2: HDAC1 (D5C6U) XP<sup>®</sup> Rabbit mAb
CUT&RUN was performed with K562 cells and either HDAC1 (D5C6U) XP® 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 CTPS1 promoter primers, human ELP6 promoter primers, and human IQCF intron 2 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: HDAC1 (D5C6U) XP<sup>®</sup> Rabbit mAb
CUT&Tag was performed with K-562 cells and HDAC1 (D5C6U) XP® 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 CTPS1 gene.
CUT & Tag Image 1: HDAC1 (D5C6U) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) orders@cellsignal.com Support: 877-678-TECH (8324) info@cellsignal.com Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 10

Cell Signaling Technology Logo
CUT&Tag was performed with K-562 cells and HDAC1 (D5C6U) XP® 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 CTPS1 gene (lower).
CUT & Tag Image 2: HDAC1 (D5C6U) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) orders@cellsignal.com Support: 877-678-TECH (8324) info@cellsignal.com Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.