Abcam, ab235634, Protein Synthesis Assay Kit (Red)

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Protein Synthesis Assay Kit (Red) ab235634 labels cells undergoing translation with a fluorescent red dye, using a method based on an alkyne-containing, cell-permeable analog of puromycin, O-Propargyl-puromycin (OP-puro). Readout on flow cytometer or fluorescence microscope.
Key facts
Detection method:Fluorescent,
Sample types:Suspension cells, Adherent cells,
Assay Platform:Flow cytometer, Fluorescence microscope

Product details:
Protein Synthesis Assay Kit (Red) (ab235634) utilizes a novel and robust chemical method based on an alkyne-containing, cell-permeable analog of puromycin, O-Propargyl-puromycin (OP-puromycin).
Protein synthesis assay principle
In the protein synthesis assay, cultured cells are incubated with OP-puromycin. Once inside the cell, OP-puromycin stops translation by forming covalent conjugates with nascent polypeptide chains.
After incubation, cells are then fixed with formalin, washed, and permeabilized with detergent.
Truncated polypeptides bound to OP-puromycin are then labelled with a fluorescent dye, in a copper-dependent reaction using a fluorescent-azide.
Cycloheximide, an inhibitor of translation, is used as a negative control. Sytox Green is used as a nuclear counter-stain.
Unlike methionine analogs, OP-puromycin does not require methionine-free conditions and can be used to label nascent proteins directly in the cell culture.
Other protein synthesis assay kits
This kit ab235634 stains cells that have undergone protein synthesis with 5(6)-TAMRA (Rhodamine), which has a red fluorescence (Ex//Em 540/580 nm). It is designed for analysis by fluorescent microscopy or flow cytometry.
Protein Synthesis Assay Kit (Green)
ab239725
stains cells that have undergone protein synthesis with 5(6)-FAM (Fluorescein), which has a green fluorescence. It is designed for analysis by fluorescent microscopy or flow cytometry.
Protein Synthesis Assay Kit (Green, Microplate)
ab273286
is an alternative version that is designed for use with a fluorometric plate reader.
Other Notes
This product was previously called K715 Biovision EZClick™ Global Protein Synthesis Assay Kit (FACS/Microscopy), Red Fluorescence. Biovision was acquired by Abcam in 2021.
Cells generate a complete set of proteins during division. Protein synthesis is a tightly regulated process and many critical controls in gene expression occur at the level of translation to ensure that production of specific cellular proteins is quickly turned on/off under specific conditions (heat chock, starvation, etc.). Protein synthesis is essential in cell growth, proliferation, signaling, differentiation or death; therefore, the identity and amount of the synthesized proteins are critical parameters in determining the physiological state of the cell. Methods enabling detection and characterization of nascent proteins, or changes in spatial and temporal protein expression/degradation patterns during disease, drug treatment or environmental changes are important tools in assesment of cytotoxicity.
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.
REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C