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.
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- PLoS genetics 19:e10110802023CRISPR loss of function screening to identify genes involved in human primordial germ cell-like cell development.Applications:Unspecified applicationReactive species:Unspecified reactive speciesYoung Sun Hwang,Yasunari Seita,M Andrés Blanco,Kotaro SasakiPubMed 38091369
- Journal of cell science 135:2022Drp1 regulates transcription of ribosomal protein genes in embryonic hearts.Applications:Unspecified applicationReactive species:Unspecified reactive speciesQiancong Zhao,Shun Yan,Jin Lu,Danitra J Parker,Huiying Wu,Qianchuang Sun,David K Crossman,Shanrun Liu,Qin Wang,Hiromi Sesaki,Kasturi Mitra,Kexiang Liu,Kai JiaoPubMed 35099001
- Cell death & disease 12:8232021Increased expression of fragmented tRNA promoted neuronal necrosis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesYanyan Cao,Kai Liu,Ying Xiong,Chunyue Zhao,Lei LiuPubMed 34462418
- Blood 139:245-2552021Targeting RIOK2 ATPase activity leads to decreased protein synthesis and cell death in acute myeloid leukemia.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJan-Erik Messling,Karl Agger,Kasper L Andersen,Kristina Kromer,Hanna M Kuepper,Anders H Lund,Kristian HelinPubMed 34359076
- Nature cell biology 23:424-4362021PANDORA-seq expands the repertoire of regulatory small RNAs by overcoming RNA modifications.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJunchao Shi,Yunfang Zhang,Dongmei Tan,Xudong Zhang,Menghong Yan,Ying Zhang,Reuben Franklin,Marta Shahbazi,Kirsty Mackinlay,Shichao Liu,Bernhard Kuhle,Emma R James,Liwen Zhang,Yongcun Qu,Qiwei Zhai,Wenxin Zhao,Linlin Zhao,Changcheng Zhou,Weifeng Gu,Jernej Murn,Jingtao Guo,Douglas T Carrell,Yinsheng Wang,Xuemei Chen,Bradley R Cairns,Xiang-Lei Yang,Paul Schimmel,Magdalena Zernicka-Goetz,Sihem Cheloufi,Ying Zhang,Tong Zhou,Qi ChenPubMed 33820973
- Oncotarget 11:2512-25302020Inhibition of the RUNX1-CBFβ transcription factor complex compromises mammary epithelial cell identity: a phenotype potentially stabilized by mitotic gene bookmarking.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJoshua T Rose,Eliana Moskovitz,Joseph R Boyd,Jonathan A Gordon,Nicole A Bouffard,Andrew J Fritz,Anuradha Illendula,John H Bushweller,Jane B Lian,Janet L Stein,Sayyed K Zaidi,Gary S SteinPubMed 32655837
- Molecules (Basel, Switzerland) 24:2019Cytostatic and Cytotoxic Natural Products against Cancer Cell Models.Applications:Unspecified applicationReactive species:Unspecified reactive speciesTaotao Ling,Walter H Lang,Julie Maier,Marizza Quintana Centurion,Fatima RivasPubMed 31130671
- Frontiers in pharmacology 10:1152019Identification of Biologically Active Compounds and Synthesis of Improved Derivatives That Confer Anti-cancer Activities .Applications:Unspecified applicationReactive species:Unspecified reactive speciesMichelle M Martínez-Montemayor,Taotao Ling,Ivette J Suárez-Arroyo,Gabriela Ortiz-Soto,Camille L Santiago-Negrón,Mercedes Y Lacourt-Ventura,Anibal Valentín-Acevedo,Walter H Lang,Fatima RivasPubMed 30837881
- Cancers 10:2018HIV-1 Envelope Protein gp120 Promotes Proliferation and the Activation of Glycolysis in Glioma Cell.Applications:Unspecified applicationReactive species:Unspecified reactive speciesGabriel Valentín-Guillama,Sheila López,Yuriy V Kucheryavykh,Nataliya E Chorna,Jose Pérez,Jescelica Ortiz-Rivera,Michael Inyushin,Vladimir Makarov,Aníbal Valentín-Acevedo,Alfredo Quinones-Hinojosa,Nawal Boukli,Lilia Y KucheryavykhPubMed 30200472
Key facts
- Detection method
- Fluorescent
- Sample types
- Suspension cells, Adherent cells
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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
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- -20°C
- Appropriate long-term storage conditions
- -20°C
- Storage information
- -20°C
Notes
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) 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) 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.
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3 product images
Functional Studies - Protein Synthesis Assay Kit (Red) (ab235634)
Analysis of protein biosynthesis in presence of Cycloheximide.
Jurkat cells (1X106 cells/mL) were pre-incubated either with vehicle or 1X Cycloheximide for 30 minutes at 37°C incubator to suppress protein synthesis followed media exchange. Cells were then incubated either with culture medium alone (white) or 1X Protein Label (red) or 1X Cycloheximide (green) for 30 minutes in the cell culture incubator. The cells were processed for protein synthesis detection by FACS according to the included protocol. Fluorescence measured in FL-2 channel clearly shows the inhibitory effect of Cycloheximide on nascent poppypeptides synthesis.
Functional Studies - Protein Synthesis Assay Kit (Red) (ab235634)
Inhibitory effect of Cycloheximide on nascent polypeptides synthesis.
Red fluorescence (upper panel) corresponds to de novo synthesized polypeptides whereas bottom panel shows the inhibitory effect of Cycloheximide on protein biosynthesis. Nuclear staining in both panels confirms that red signal is a result of Protein Label incorporation.
Functional Studies - Protein Synthesis Assay Kit (Red) (ab235634)
FACS analysis of the inhibitory effect of Cycloheximide.
FACS analysis of negative control (white), positive control (Protein Label, red) and Cycloheximide-treated (green) cell populations. Signal measured in FL-2 channel clearly shows the inhibitory effect of Cycloheximide on nascent polypeptides synthesis.
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