Product name

MitoPT TMRM Mitochondrial Depolarization Assay Kit

Full Product Name

MitoPT TMRM Mitochondrial Depolarization Assay Kit

Product gene name

MitoPT TMRM Assay Kit

Reagents: TMRM

Sample protocol

Cell culture up to 1 x 10 ^ 6 cells / mL. Expose cells to an apoptosis-inducing agent or metabolic stress environment using your experimental protocol. Reconstitute the TMRM reagent with 100 uL DMSO to form the stock concentrate (can be frozen for future use). Dilute the stock concentrate with 1X PBS, cell culture medium, or wash buffer to achieve an intermediate stock of 10 uM to add aliquots of cells.

Pour the cell cultures with 10 uM TMRM to achieve a TMRM concentration of 20-200 nM. Incubate cells in TMRM solution (20-200 nM), 15-20 minutes at 37 degrees C in the dark. Wash cells once in wash buffer if using TMRM staining concentration> 50 nM. Analyze the data with a fluorescence microscope, plate reader, or flow cytometer.

Objective: Mitochondrial depolarization

Excitation / Emission: 548 nm / 573 nm

Analysis method

Flow cytometer, fluorescence microscope, fluorescence plate reader

Sample types: Cell culture, tissue

Preparation and storage

• Mixed storage conditions: CCCP at -20 degrees C
• Other kit components at 2-8 degrees C

Product note

Information selected from the online data sheet is extracted from bioinformatics databases, occasionally resulting in the ambiguous or irrelevant product information. It is the customer’s responsibility to review, verify and evaluate the information to make sure it matches their requirements before purchasing the kit. The essays in our ELISA kit are dynamic research tools and can sometimes be updated and improved.

If the format of this assay is important to you, please request the current manual or contact our technical support team with a pre-sale consultation before placing an order. We will confirm the current details of the trial. We cannot guarantee that the sample manual posted online is the most current manual, it is intended to serve as an example only.

Other notes

Occasionally, small volumes of MitoPT TMRM Assay Kit vials may become trapped in the product vial seal during shipping and storage. If necessary, briefly centrifuge the vial in a benchtop centrifuge to dislodge any liquid on the lid of the container. Some products may need to be shipped with dry ice and an additional fee may apply for dry ice.

Related Product Information for MitoPT TMRM Test Kit

• Description:

MitoPT TMRM Assay Kits use the well-characterized lipophilic but slightly cationic TMRM fluorescent dye to detect mitochondrial membrane permeability. The use of the MitoPT TMRM kit allows us to easily distinguish between apoptotic cells or cells compromised with the mitochondrial membrane and non-apoptotic cells with polarized mitochondria. Cells with polarized mitochondria will emit bright red fluorescence (max em: 573 nm); cells with cells committed to the membrane will exhibit attenuated fluorescence or no signal.

The MitoPT TMRM mitochondrial membrane permeability assay can be used in conjunction with existing apoptosis or metabolic stress protocols. Each kit includes the TMRM potentiometric dye reagent, a 10X wash buffer to remove any excess TMRM dye after staining, and the generic mitochondrial membrane depolarizer, carbonylcyanide m-chlorophenylhydrazone (CCCP). TMRM-stained cells can be analyzed using flow cytometry, fluorescence microscopy, and fluorescence plate reader evaluation methodology.

• Background:

Detecting Mitochondrial Permeability Transition and Mitochondrial Membrane Depolarization with the MitoPT TMRM Assay The ICT MitoPT TMRM Assay uses the well-characterized fluorescent potentiometric tetramethylrhodamine methyl ester (TMRM) to detect mitochondrial permeability and depolarization of the membrane in whole living cells. The rhodamine-based TMRE and TMRM dyes offered in ICT MitoPT cell-based assay kits are classified as slow membrane redistributing dyes. The lipophilic structures of these dyes allow them to easily penetrate membrane barriers.

Precautions

All MyBioSource products are for scientific laboratory research purposes and not for diagnostic, therapeutic, prophylactic or in vivo use. Through your purchase, you expressly represent and warrant to MyBioSource that you will properly test and use any Product purchased from MyBioSource in accordance with industry standards. MyBioSource and its authorized distributors reserve the right to refuse to process any order when we reasonably believe that the intended use will be outside of our acceptable guidelines.

Disclaimer

While every effort has been made to ensure the accuracy of the information provided in this datasheet, MyBioSource will not be responsible for any omissions or errors contained herein. MyBioSource reserves the right to make changes to this datasheet at any time without prior notice.