Immunohistochemistry was performed on normal biopsies of deparaffinized human skin tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a Rabbit Polyclonal Antibody recognizing HDAC4 (PA1-863 ) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues M(1)SSQSHPDGLSGRDQPVE(18) of human HDAC4|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||1-3 ul|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20|
|Western Blot (WB)||1:1,000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
PA1-863 detects histone deacetylase 4 (HDAC4) from human cells.
PA1-863 has been successfully used in Western blot and ChIP procedures. By Western blot, this antibody detects an ~140 kDa protein representing HDAC4 from HeLa cell lysate. An ~110 kDa protein is also detected with PA1-863 which could correspond to protein degradation product.
PA1-863 immunogen is a synthetic peptide corresponding to residues M(1)SSQSHPDGLSGRDQPVE(18) of human HDAC4.
Chromatin is a highly specialized structure composed of tightly compacted chromosomal DNA. Gene expression within the nucleus is controlled, in part, by a host of protein complexes which continuously pack and unpack the chromosomal DNA. One of the known mechanisms of this packing and unpacking process involves the acetylation and deacetylation of the histone proteins comprising the nucleosomal core. Acetylated histone proteins confer accessibility of the DNA template to the transcriptional machinery for expression. Histone deacetylases (HDACs) are chromatin remodeling factors that deacetylate histone proteins and thus, may act as transcriptional repressors. HDACs are classified by their sequence homology to the yeast HDACs and there are currently 2 classes. Class I proteins are related to Rpd3 and members of class II resemble Hda1p.
HDAC4 is a class II histone deacetylase containing 1084 amino acid residues. HDAC4 has been shown to interact with N-CoR.
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