Revision 1

#65816

Store at -20C

Class I HDAC Antibody Sampler Kit

1 Kit

(3 x 20 microliters)

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.

Product Includes Product # Quantity Mol. Wt Isotype/Source
HDAC1 (D5C6U) Rabbit Monoclonal Antibody3458920 µl62 kDaRabbit IgG
HDAC2 (D6S5P) Rabbit Monoclonal Antibody5715620 µl60 kDaRabbit IgG
HDAC3 (D2O1K) Rabbit Monoclonal Antibody8505720 µl49 kDaRabbit IgG
Anti-rabbit IgG, HRP-linked Antibody7074100 µlGoat

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The Class I HDAC Antibody Sampler Kit provides an economical means of detecting Class I HDAC proteins using control antibodies against HDAC1, HDAC2 and HDAC3. The kit contains enough primary antibodies to perform at least two western blot experiments.

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.

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).

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 1

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using HDAC1 (D5C6U) XP® Rabbit mAb.
undefined Image 1: Class I HDAC Antibody Sampler Kit
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.
undefined Image 2: Class I HDAC Antibody Sampler Kit
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).
undefined Image 3: Class I HDAC Antibody Sampler Kit
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 1

Cell Signaling Technology Logo

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

undefined Image 4: Class I HDAC Antibody Sampler Kit
CUT&RUN was performed with K562 cells and either HDAC2 (D6S5P) Rabbit Monoclonal Antibody or Phospho-HDAC2 (Ser394) (E8O2Z) Rabbit Monoclonal Antibody #69238, 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 HDAC2 (see additional figure containing CUT&RUN-qPCR data).
undefined Image 5: Class I HDAC Antibody Sampler Kit
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
undefined Image 6: Class I HDAC Antibody Sampler Kit
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 1

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using HDAC3 (D2O1K) Rabbit mAb.
undefined Image 7: Class I HDAC Antibody Sampler Kit
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.
undefined Image 8: Class I HDAC Antibody Sampler Kit
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).
undefined Image 9: Class I HDAC Antibody Sampler Kit
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 1

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).
undefined Image 10: Class I HDAC Antibody Sampler Kit
CUT&RUN was performed with K562 cells and either HDAC2 (D6S5P) Rabbit Monoclonal Antibody or Phospho-HDAC2 (Ser394) (E8O2Z) Rabbit Monoclonal Antibody #69238, 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 HDAC2 (see additional figure containing CUT&RUN-qPCR data).
undefined Image 11: Class I HDAC Antibody Sampler Kit
Chromatin immunoprecipitations were performed with cross-linked chromatin from MCF7 cells treated with tamoxifen (100 nM, 45 min) and HDAC3 (D2O1K) 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. The figure shows binding across TFF1/pS2, a known target gene of HDAC3 (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.
undefined Image 12: Class I HDAC Antibody Sampler Kit
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 1

Cell Signaling Technology Logo
Confocal immunofluorescent analysis of HeLa cells using HDAC1 (D5C6U) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red).
undefined Image 13: Class I HDAC Antibody Sampler Kit
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.
undefined Image 14: Class I HDAC Antibody Sampler Kit

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

undefined Image 15: Class I HDAC Antibody Sampler Kit
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 1

Cell Signaling Technology Logo
CUT&RUN was performed with K562 cells and either HDAC2 (D6S5P) Rabbit Monoclonal Antibody or Rabbit (DA1E) Monoclonal Antibody IgG 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.
undefined Image 16: Class I HDAC Antibody Sampler Kit
Chromatin immunoprecipitations were performed with cross-linked chromatin from MCF7 cells treated with tamoxifen (100 nM, 45 min) and HDAC3 (D2O1K) 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. The figure shows binding across chromosome 21 (upper), including TFF1/pS2 (lower), a known target gene of HDAC3 (see additional figure containing ChIP-qPCR data).
undefined Image 17: Class I HDAC Antibody Sampler Kit
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.
undefined Image 18: Class I HDAC Antibody Sampler Kit
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 1

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using HDAC1 (D5C6U) XP® Rabbit mAb.
undefined Image 19: Class I HDAC Antibody Sampler Kit
Chromatin immunoprecipitations were performed with cross-linked chromatin from MCF7 cells treated with tamoxifen (100 nM, 45 min) and either HDAC3 (D2O1K) 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 pS2 Promoter Primers #9702, SimpleChIP® Human c-Myc Intron 1 Primers #14905, human c-Myc promoter primers, 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.
undefined Image 20: Class I HDAC Antibody Sampler Kit
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).
undefined Image 21: Class I HDAC Antibody Sampler Kit
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 1

Cell Signaling Technology Logo

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

undefined Image 22: Class I HDAC Antibody Sampler Kit
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.
undefined Image 23: Class I HDAC Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human urothelial carcinoma using HDAC1 (D5C6U) XP® Rabbit mAb.
undefined Image 24: Class I HDAC Antibody Sampler Kit
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 1

Cell Signaling Technology Logo

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

undefined Image 25: Class I HDAC Antibody Sampler Kit

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

undefined Image 26: Class I HDAC Antibody Sampler Kit
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.
undefined Image 27: Class I HDAC Antibody Sampler Kit
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 1

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded CT26.WT syngeneic tumor using HDAC1 (D5C6U) XP® Rabbit mAb.
undefined Image 28: Class I HDAC Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded Renca syngeneic tumor using HDAC1 (D5C6U) XP® Rabbit mAb.
undefined Image 29: Class I HDAC Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded mouse brain using HDAC1 (D5C6U) XP® Rabbit mAb.
undefined Image 30: Class I HDAC Antibody Sampler Kit
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 1

Cell Signaling Technology Logo

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

undefined Image 31: Class I HDAC Antibody Sampler Kit

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

undefined Image 32: Class I HDAC Antibody Sampler Kit
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). 
undefined Image 33: Class I HDAC Antibody Sampler Kit
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 1

Cell Signaling Technology Logo
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.
undefined Image 34: Class I HDAC Antibody Sampler Kit
Western blot analysis of extracts from various cell lines using HDAC2 (D6S5P) Rabbit mAb.
undefined Image 35: Class I HDAC Antibody Sampler Kit
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).
undefined Image 36: Class I HDAC Antibody Sampler Kit
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 1

Cell Signaling Technology Logo
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.
undefined Image 37: Class I HDAC Antibody Sampler Kit
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.
undefined Image 38: Class I HDAC Antibody Sampler Kit
Confocal immunofluorescent analysis of HeLa cells using HDAC2 (D6S5P) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red).
undefined Image 39: Class I HDAC Antibody Sampler Kit
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 1

Cell Signaling Technology Logo
Immunoprecipitation of HDAC3 protein from MCF-7 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is HDAC3 (D2O1K) Rabbit mAb. Western blot analysis was performed using HDAC3 (D2O1K) Rabbit mAb #85057. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as a secondary antibody.
undefined Image 40: Class I HDAC Antibody Sampler Kit
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.