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65816
Class I HDAC Antibody Sampler Kit

Class I HDAC Antibody Sampler Kit #65816

Western Blotting Image 1

Western blot analysis of extracts from various cell lines using HDAC1 (D5C6U) XP® Rabbit mAb.

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Western Blotting Image 2

Western blot analysis of extracts from various cell lines using HDAC2 (D6S5P) Rabbit mAb.

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Western Blotting Image 3

Western blot analysis of extracts from various cell lines using HDAC3 (D2O1K) Rabbit mAb.

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Western Blotting Image 4

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.

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IP Image 5

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.

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Chromatin IP Image 6

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 K-562 cells and either 10 μl of HDAC1 (D5C6U) XP® Rabbit mAb or 2 μl of 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.

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IP 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.

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Chromatin IP Image 8

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 K562 cells and either 10 µl of HDAC2 (D6S5P) Rabbit mAb or 2 µl of 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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Chromatin IP Image 9

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 MCF7 cells treated with tamoxifen (100 nM, 45 min) and either 10 μl of HDAC3 (D2O1K) Rabbit mAb or 2 μl of 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.

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IF-IC Image 10

Confocal immunofluorescent analysis of HeLa cells using HDAC1 (D5C6U) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red).

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IF-IC Image 11

Confocal immunofluorescent analysis of HeLa cells using HDAC2 (D6S5P) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red).

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
HDAC1 (D5C6U) XP® Rabbit mAb 34589 20 µl
  • WB
  • IP
  • IF
  • ChIP
H M R Mk 62 Rabbit IgG
HDAC2 (D6S5P) Rabbit mAb 57156 20 µl
  • WB
  • IP
  • IF
  • ChIP
H M R Mk 60 Rabbit IgG
HDAC3 (D2O1K) Rabbit mAb 85057 20 µl
  • WB
  • IP
  • ChIP
H M R Mk 49 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

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.

Each antibody in the Class I HDAC Antibody Sampler Kit detects endogenous levels of its target protein. Cross-reactivity was not observed with other family members.

Monoclonal antibodies are produced by immunizing rabbits with synthetic peptides corresponding to Ala440 of human HDAC1, His438 of human HDAC3, and Gly416 of human HDAC3 proteins.

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.
Entrez-Gene Id
3065 , 3066 , 8841
Swiss-Prot Acc.
Q13547 , Q92769 , O15379
For Research Use Only. Not For Use In Diagnostic Procedures.

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

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