Home >Protein Extraction & Purification >Mammalian Cell & Tissue Extraction Kit

Description

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The Mammalian Cell & Tissue Extraction Kit provides optimized cell extraction buffer, protease inhibitor cocktail, and DTT for convenient extraction of mammalian proteins from cultured cells and tissue samples, under nondenaturing conditions. Cell lysate prepared using the kit can be used in a variety of applications, such as enzyme activity assays (e.g., caspase activity assays), Western blot analysis, and others. The entire procedure takes less than 20 minutes.

Datasheet

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Cat # +Size	K269-500
Size	500 assays
Species Reactivity	Mammalian
Applications	Applications- Enzyme activity assays (e.g., caspase activity assays), Western blot analysis, and others.
Features & Benefits	• Simple procedure; takes less than 20 minutes • Fast and convenient • The Mammalian Cell Extraction Kit provides optimized lysis buffer and protease inhibitor cocktail as well as other reagents for extraction of mammalian proteins from tissues and cells under nondenaturing conditions.
Kit Components	• Cell Extraction Buffer • Protease Inhibitor Cocktail • DTT (1 M)
Storage Conditions	-20°C
Shipping Conditions	Gel Pack
USAGE	For Research Use Only! Not For Use in Humans.

FAQ

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How to estimate the protein? Can we use a BCA assay?
You can use a reducing agent compatible BCA assay.

What is the difference between Extraction and lysis buffer?
Extraction buffer is meant to gently loosen the plasma membrane and leak out the contents of the cell unlike a lysis buffer. It is non-denaturing to help preserve function of all the proteins isolated which also makes it gentler than a conventional denaturing lysis buffer. Total mg of protein extracted might be less than a conventional lysis buffer but our solution makes sure that all proteins retain biological function and native structure.

Does this kit extract whole cell protein?
The extraction buffer helps lyse the plasma membrane primarily. During the top-speed spin the nuclei and broken plasma membrane debris are pelleted down and only the cytosol along with the ER, Golgi etc remain in the lysate. Typically nuclei are not broken by this lysis process.

Does the lysis buffer break open the nuclei to release proteins?
"The extraction buffer helps lyse the plasma membrane primarily. During the top-speed spin the nuclei and broken plasma membrane debris are pelleted down and only the cytosol along with the ER, Golgi etc remain in the lysate. Typically nuclei are not broken by this lysis process."

Can the kit work on bacteria or yeast cells?
The kit has been standardized for mammalian cells only.

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.

CITATIONS

1. Juan Li, Autophagy Induction by HIV-Tat and Methamphetamine in Primary Midbrain Neuronal Cells of Tree Shrews via the mTOR Signaling and ATG5/ATG7 Pathway. Front Neurosci., Dec 2018; 30574066.
2. Chunyan Gu, Upregulation of FOXM1 leads to diminished drug sensitivity in myeloma. BMC Cancer, Nov 2018; 30463534.
3. Gu Z et al., (2017) NEK2 Promotes Aerobic Glycolysis in Multiple Myeloma Through Regulating Splicing of Pyruvate Kinase, J Hematol Oncol, 2017, 13;10(1):17
4. Cosenza, Takashi et al. (2017) Ricolinostat, a selective HDAC6 inhibitor, shows anti-lymphoma cell activity alone and in combination with bendamustine, Apoptosis. 2017 Jun;22(6):827-840.
5. Shen, Cunzhou et al. (2016) Potential protective effects of autophagy activated in MPP+ treated astrocytes, Exp Ther Med. 2016 Nov;12(5):2803-2810.

Downloads

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Datasheet (pdf)
Safety Data Sheets (SDS) (pdf)