Revision 1

#65816Store at -20C

1 Kit

(3 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
HDAC1 (D5C6U) XP® Rabbit mAb 34589 20 µl 62 kDa Rabbit IgG
HDAC2 (D6S5P) Rabbit mAb 57156 20 µl 60 kDa Rabbit IgG
HDAC3 (D2O1K) Rabbit mAb 85057 20 µl 49 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

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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).

  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.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a 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.

    使用に関する制限

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    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
    #65816

    Class I HDAC Antibody Sampler Kit

    Class I HDAC Antibody Sampler Kit: Image 1 Expand Image
    Western blot analysis of extracts from various cell lines using HDAC1 (D5C6U) XP® Rabbit mAb.
    Class I HDAC Antibody Sampler Kit: Image 2 Expand Image
    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).
    Class I HDAC Antibody Sampler Kit: Image 3 Expand Image
    Western blot analysis of extracts from various cell lines using HDAC2 (D6S5P) Rabbit mAb.
    Class I HDAC Antibody Sampler Kit: Image 4 Expand Image
    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.
    Class I HDAC Antibody Sampler Kit: Image 5 Expand Image
    Western blot analysis of extracts from various cell lines using HDAC3 (D2O1K) Rabbit mAb.
    Class I HDAC Antibody Sampler Kit: Image 6 Expand Image
    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.
    Class I HDAC Antibody Sampler Kit: Image 7 Expand Image
    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).
    Class I HDAC Antibody Sampler Kit: Image 8 Expand Image
    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.
    Class I HDAC Antibody Sampler Kit: Image 9 Expand Image
    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.
    Class I HDAC Antibody Sampler Kit: Image 10 Expand Image
    Confocal immunofluorescent analysis of HeLa cells using HDAC1 (D5C6U) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red).
    Class I HDAC Antibody Sampler Kit: Image 11 Expand Image
    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.
    Class I HDAC Antibody Sampler Kit: Image 12 Expand Image
    Confocal immunofluorescent analysis of HeLa cells using HDAC2 (D6S5P) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red).
    Class I HDAC Antibody Sampler Kit: Image 13 Expand Image
    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).
    Class I HDAC Antibody Sampler Kit: Image 14 Expand Image
    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.
    Class I HDAC Antibody Sampler Kit: Image 15 Expand Image
    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.
    Class I HDAC Antibody Sampler Kit: Image 16 Expand Image
    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.
    Class I HDAC Antibody Sampler Kit: Image 17 Expand Image
    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).
    Class I HDAC Antibody Sampler Kit: Image 18 Expand Image
    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).
    Class I HDAC Antibody Sampler Kit: Image 19 Expand Image
    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.
    Class I HDAC Antibody Sampler Kit: Image 20 Expand Image
    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.
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    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.