Human HDAC4 Baculovirus-Insect cells Overexpression Lysate: Product Information
This Human HDAC4 overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of HDAC4 protein (Cat: 11807-HNCB) from the overexpression lysate was verified.
A DNA sequence encoding the human HDAC4 (NP_006028.2)(Met612-Leu1084) was expressed and purified with two additional amino acids (Gly & Pro ) at the N-terminus.
The secreted recombinant human HDAC4 consists of 475 amino acids and predicts a molecular mass of 50.9 KDa. The apparent molecular mass of the protein is approximately 51 KDa in SDS-PAGE under reducing conditions due to glycosylation.
Human HDAC4 Baculovirus-Insect cells Overexpression Lysate: Usage Guide
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube.
2. Re-dissolve the pellet using 200μL pure water and boil for 2-5 min.
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Stability & Storage
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
Western Blot (WB) Optimal dilutions/concentrations should be determined by the end user.
Human HDAC4 Baculovirus-Insect cells Overexpression Lysate: Synonyms
Human AHO3 Overexpression Lysate; Human BDMR Overexpression Lysate; Human HA6116 Overexpression Lysate; Human HD4 Overexpression Lysate; Human HDAC-4 Overexpression Lysate; Human HDAC-A Overexpression Lysate; Human HDACA Overexpression Lysate
HDAC4 Background Information
HDAC4 (histone deacetylase 4), belongs to class II of the histone deacetylase/AcuC/APhA family. Histone Deacetylases (HDACs) are a group of enzymes closely related to sirtuins. They catalyze the removal of acetyl groups from lysine residues in histones and non-histone proteins, resulting in transcriptional repression. In general, they do not act autonomously but as components of large multiprotein complexes, such as pRb-E2F and mSin3A, that mediate important transcription regulatory pathways. There are three classes of HDACs; classes 1, 2, and 4, which are closely related to Zn2+-dependent enzymes. HDACs are ubiquitously expressed and they can exist in the nucleus or cytosol. Their subcellular localization is affected by protein-protein interactions and by the class to which they belong. HDACs have a role in cell growth arrest, differentiation, and death and this has led to substantial interest in HDAC inhibitors as possible antineoplastic agents. HDAC4 possesses histone deacetylase activity and represses transcription when tethered to a promoter. It does not bind DNA directly but through transcription factors MEF2C and MEF2D. HDAC4 seems to interact in a multiprotein complex with RbAp48 and HDAC3.
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