Confocal immunofluorescent analysis of HeLa cells using HDAC2 (3F3) Mouse mAb (green). Actin filaments were labeled using DY-554 Phalloidin (red).
Confocal immunofluorescent analysis of HeLa cells using HDAC3 (7G6C5) Mouse mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).
Western blot analysis of extracts from various cell lines using HDAC4 (D15C3) Rabbit mAb.
Western blot analysis of NIH/3T3 cells using Phospho-HDAC4 (Ser246)/HDAC5 (Ser259)/HDAC7 (Ser155) (D27B5)
Rabbit mAb #3443, HDAC4 (D8T3Q) Rabbit mAb #15164, HDAC5 (D1J7V) Rabbit mAb #20458, and HDAC7 (D4E1L) Rabbit mAb #33418. To show the phospho-specificity of the antibody, the blot was mock treated (-) or treated (+) with calf intestinal phosphatase (CIP) and lambda phosphatase. As expected, signal from the Phospho-HDAC4 (Ser246)/HDAC5 (Ser259)/HDAC7 (Ser155) (D27B5) Rabbit mAb is lost after treatment of the blot with phosphatase.
Flow cytometric analysis of K562 cells using HDAC6 (D2E5) Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
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.
Western blot analysis of extracts from HeLa, NIH/3T3, and H-4-II-E cells using HDAC1 (10E2) Mouse mAb.
Western blot analysis of extracts from various cell lines using HDAC2 (3F3) Mouse mAb.
Western blot analysis of extracts from various cell lines using HDAC3 (7G6C5) Mouse mAb.
Confocal immunofluorescent analysis of A549 cells, untreated (left) or treated with MG132 (5 μM, 24 hr; right), using HDAC6 (D2E5) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5®#4084 (fluorescent DNA dye).
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using HDAC6 (D2E5) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using HDAC6 (D2E5) Rabbit mAb.
Western blot analysis of extracts from various cell lines using HDAC6 (D2E5) Rabbit mAb.
|HDAC1 (10E2) Mouse mAb 5356||20 µl||
||H Mk M R||62||Mouse IgG1|
|HDAC2 (3F3) Mouse mAb 5113||20 µl||
||H Mk M R||60||Mouse IgG1|
|HDAC3 (7G6C5) Mouse mAb 3949||20 µl||
||H Mk M R||49||Mouse IgG2a|
|HDAC4 (D15C3) Rabbit mAb 7628||20 µl||
||H Mk M R||140||Rabbit IgG|
|Phospho-HDAC4 (Ser246)/HDAC5 (Ser259)/HDAC7 (Ser155) (D27B5) Rabbit mAb 3443||20 µl||
||H M||140, 124||Rabbit IgG|
|HDAC6 (D2E5) Rabbit mAb 7558||20 µl||
||H Mk||160||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody 7074||100 µl||
|Anti-mouse IgG, HRP-linked Antibody 7076||100 µl||
The Histone Deacetylase (HDAC) Antibody Sampler Kit provides a fast and economical means to evaluate the endogenous levels of HDACs. The kit contains enough primary and secondary antibodies to perform two Western blot experiments.
The Histone Deacetylase (HDAC) Antibody Sampler Kit provides a fast and economical means to evaluate the endogenous levels of HDACs. The kit contains enough primary and secondary antibodies to perform four mini-blot experiments.
Each antibody in the Histone Deacetylase (HDAC) Antibody Sampler Kit detects endogenous levels of its target.
Monoclonal antibodies are produced by immunizing animals with a recombinant protein specific to the carboxy terminus of human HDAC6, the amino terminus of human HDAC4 protein, synthetic peptides to the carboxy-terminal residues of human HDAC1 and HDAC2 proteins, or synthetic phosphopeptide corresponding to Ser155 of human HDAC7 protein. HDAC3 (7G6C5) monoclonal antibody is produced by immunizing animals with recombinant human HDAC3 protein.
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).
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