Histones are small alkaline proteins that interact with eukaryotic DNA to form the fundamental subunit of chromatin known as the nucleosome. The core histones H2A, H2B, H3 and H4 form the central spooling unit of the nucleosome; with two copies of each comprising a histone octamer that interacts with the negatively charged DNA. They enable an ordered compaction of the DNA, as well as spatiotemporal control of gene expression and genomic replication throughout mitosis. Post-translational modification of this core complex results in structural changes within the chromatin and epigenetic control of gene expression. BioVision’s EZExtract™ Core Histone Isolation Kit offers an optimized suite of reagents to rapidly extract the total core histones while preserving acetylation from tissues and cultured cells for biochemical and epigenetic analyses. One hundred standard extractions at a scale of 107 cells or 100 mg tissues can be performed with the reagents in this kit. Yield of the total histone proteins is approximately 0.4 mg per 107 cells or 100 mg of tissue
|Cat # +Size||K234-100|
|Applications||Isolation of intact functional core histones from tissues and cultured cells Protein profiling, post-translational modification and epigenetic analyses Protein profiling of Histones by SDS-PAGE and Western blot|
|Features & Benefits||• Simple & convenient protocol
• The kit provides unique formulations of ready-to-use buffers and reagents to isolate histones with minimal sample preparation
|Kit Components||• Lysis Buffer
• Extraction Reagent
• Neutralizing Buffer • DTT (1 M)
|Shipping Conditions||Gel Pack|
|USAGE||For Research Use Only! Not For Use in Humans.|
Do the histone proteins isolated using this kit also have the modifications intact?
The protocol helps in preserving all the modification on the extracted histone proteins.
Can we use frozen samples with this assay?
We recommend using fresh samples. However, frozen samples can also be used. Samples should be frozen at -80C and must not have gone through multiple freeze/thaw cycles.
Does the kit contain HDAC inhibitors?
No the kit does not contain any HDAC inhibitor.
Can we use RIPA buffer to isolate histone proteins as opposed to K234-100?
This kit is specifically helpful in isolating histone proteins. The isolated histone protein concentration is much higher with this kit compared to simple RIPA buffer.
Does the purified product need to be de-salted before running on a gel?
No special preparation such as desalting needs to be done before loading the sample into the gel. If salt precipitates are seen in the extracts after being frozen, thaw extracts on ice and pipette gently several times until salts are re-dissolved.
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 yopur regional BioVision distributor.
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.
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Our products such as Elisa, Antibodies, Proteins, Peptides and sequencing kits are covered by Biolinkk quality warranty and will work as described in datasheet, a free replacement or money back is guaranteed if does not perform according to datasheet.