Histone deacetylases (HDACs) play a central role in controlling cell cycle regulation, cell differentiation, and tissue development. HDAC1 is ubiquitously expressed and plays a role in cellular senescence, cardiomyocyte differentiation and hypertrophy, aging of the liver, myelination, and adult neurogenesis to name a few functions. The HDAC1 IP & Activity Assay Kit provides an antibody-based method to specifically immunoprecipitate the HDAC1 complex from cells & tissues and to measure HDAC1 activity fluorometrically. HDAC1 is immunoprecipitated from cell, nuclear or tissue extract(s) using HDAC1 antibody followed by capturing the complex on protein-A/G beads. The immunoprecipitated complex is incubated with the HDAC substrate. Only the deacetylated substrate is cleaved by the Developer to produce a fluorophore, which can be easily analyzed using a fluorescence plate reader.
|Cat # +Size||K341-25|
|Detection Method||Fluorescence (Ex/Em=380/500 nm)|
|Applications||Immunoprecipitation of HDAC2 complex from cell, nuclear, or tissue extract(s) Measurement of HDAC2 activity of immunoprecipitated complex/purified enzyme Screening for activators or inhibitors of HDAC2|
|Features & Benefits||• Simple, fast and convenient.
• Only kit on the market for both IP as well as measuring activity of HDAC2
|Kit Components||• HDAC Assay Buffer|
• Cell Lysis Buffer
• HDAC Substrate
• AFC (7-amino-4-trifluoromethyl coumarin) Standard (1 mM)
• Rabbit HDAC2 Antibody (0.2 mg/ml)
• Rabbit IgG (Control Antibody) (0.5 mg/ml)
• Protein A/G Sepharose Beads
• Positive Control (Jurkat Cell Lysate) (125 μg)
|Shipping Conditions||Gel Pack|
|USAGE||For Research Use Only! Not For Use in Humans.|
Does the kit detect the total activity of HDAC or the activity of HDAC 2?
The HDAC2 antibody in K341 is a high affinity polyclonal antibody that recognizes multiple epitopes on the HDAC2 protein. Hence it should only specifically pull down HDAC2. The substrate is an HDAC substrate, not a HDAC-2 specific substrate. There is always overlap in the acetylation ability of HDACs. So the point of quality control is the antibody pull-down.
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact your regional BioVision distributor.
What is the shelf life of this kit?
BioVisions’s assay kits expire 6 months from the date of shipment. Some components of the kit may expire sooner as mentioned in the data sheet
Can we purchase individual components of this kit?
Yes, you can purchase any of the kit's components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.
Kazunori Inoue, Podocyte histone deacetylase activity regulates murine and human glomerular diseases. J Clin Inves, Feb 2019; 30776024.
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