Cell Based Assay

α-Ketoglutarate (α-KG) is an important intermediate of the TCA cycle. It is used as an indicator of enhanced uptake of glucose metabolites into the TCA cycle and of enhanced glutaminolysis, the metabolic pathway for supplying α-ketoglutarate using glutamine as a substrate. α-Ketoglutarate plays an essential role in the production of glutamate and γ-aminobutyric acid (GABA) and contributes to the scavenging of reactive oxygen species. Additionally, a recent cancer study suggested that...

Detection Principle -Bacstain- Bacterial Viability Detection Kits are a series of products for fluorescent double-staining of bacteria. By combining different types of fluorescent stain, staining images can be acquired for each index (membrane impairment, respiratory activity, and esterase activity). Bacterial viability is generally assessed by colony formation in the nutrient-agar medium. However, this requires a long culturing time, and it is hard to recognize the growth of viable but...

Detection Principle -Bacstain- Bacterial Viability Detection Kits are a series of products for fluorescent double-staining of bacteria. By combining different types of fluorescent stain, staining images can be acquired for each index (membrane impairment, respiratory activity, and esterase activity). Bacterial viability is generally assessed by colony formation in the nutrient-agar medium. However, this requires a long culturing time, and it is hard to recognize the growth of viable but...

Detection Principle -Bacstain- Bacterial Viability Detection Kits are a series of products for fluorescent double-staining of bacteria. By combining different types of fluorescent stain, staining images can be acquired for each index (membrane impairment, respiratory activity, and esterase activity). Bacterial viability is generally assessed by colony formation in the nutrient-agar medium. However, this requires a long culturing time, and it is hard to recognize the growth of viable but...

Fig. 1 Cell staining mechanism

Required Equipment and Materials 10 μl, 1000 μl pipettes, incubator, Microscope (blue excitation filter and red emission filter) or flow cytometer (488 nm blue laser)Fig. 1 Cell staining mechanism ...

BS01 BS02 Product Description Colony formation using an agar plate is a very common and reliable method for counting bacterial cells. However, it takes quite a long time to form a colony. Therefore, alternative detection methods have been developed. Bacteria-specific gene amplification methods such as PCR, LAMP, and nucleus staining are quite rapid, but these methods count dead bacteria as well. Therefore, detection of live cell functions is essential to determining the actual number of living...

Chemical Structure

Chemical Structure

Product Description Acridine orange (AO) forms a complex with double-stranded DNA to emit green fluorescence (Fig. 1). AO also forms a complex with singlestranded DNA or RNA to emit red fluorescence. One molecule of AO intercalates with three base pairs of double-stranded DNA and emits green fluorescence with the maximum wavelength at 526 nm (excitation 502 nm). One molecule of AO can also interact with one phosphate group of single-stranded DNA or RNA to form an aggregated, or stacked,...

Product Description Calcein-AM readily passes through the cell membrane of viable cells because of its enhanced hydrophobicity compared to Calcein. After Calcein-AM permeates into the cytoplasm, it is hydrolyzed by esterases to Calcein, which remains inside the cell (Fig. 1). Among other reagents, including BCECF-AM and Carboxy-fluorescein diacetate, Calcein-AM is the most suitable fluorescent probe for staining viable cells because of its low cytotoxicity. Calcein does not inhibit any...

Product Description Calcein-AM readily passes through the cell membrane of viable cells because of its enhanced hydrophobicity compared to Calcein. After Calcein-AM permeates into the cytoplasm, it is hydrolyzed by esterases to Calcein, which remains inside the cell (Fig. 1). Among other reagents, including BCECF-AM and Carboxy-fluorescein diacetate, Calcein-AM is the most suitable fluorescent probe for staining viable cells because of its low cytotoxicity. Calcein does not inhibit any...

Product Description CFSE is cell-membrane permeable and readily accumulates inside viable cells where it covalently attaches to intracellular proteins (Fig. 1). Hydrolyzed CFSE emits fluorescence and covalently attached fluorescein molecules do not leak from cells. CFSE-labeled cells can be monitored over several weeks in vivo. Therefore, CFSE is utilized for detection of viable cell as well as for the long-term observation of cell activities by fluorescent microscopy. The excitation and...

Product Description CytoRed is cell membrane permeable and accumulates inside of viable cells as resorufin (Fig. 1). CytoRed has a much wider spectrum than BCECF or Calcein, so filters for fluorescein and rhodamine can also be used. The excitation and emission wavelengths of resorufin are 560 nm and 590 nm, respectively. A photo of cells stained with CytoRed is shown in Fig. 2. Fig. 1 Cell staining mechanism ...

Product Description DAPI is an AT-sequence-specific DNA intercalator that attaches to DNA at the minor groove of the double helix like Hoechst dyes. DAPI is not permeable through viable cell membranes, but it passes through disturbed cell membranes to stain the nucleus. DAPI has a high photobleaching tolerance level. DAPI is utilized for the detection of mitochondrial DNA in yeast, chloroplast DNA, virus DNA, micoplasm DNA, and chromosomal DNA. The excitation and emission wavelengths of...

Product Description DAPI is an AT-sequence specific DNA intercalator that attaches to DNA at the minor groove of the double helix like Hoechst dyes. Though DAPI is not permeable through viable cell membranes, it passes through disturbed cell membranes to stain the nucleus. DAPI has a high photo-bleaching tolerance level. DAPI is utilized for the detection of mitochondrial DNA in yeast, chloroplast DNA, virus DNA, micoplasm DNA and chromosomal DNA. The excitation and emission wavelengths of...

Product Description -Cellstain-Double Staining Kit is utilized for simultaneous fluorescence staining of viable and dead cells. This kit contains Calcein-AM and Propidium Iodide (PI) solutions, which stain viable and dead cells, respectively (Fig. 1). Calcein-AM, an acetoxymethyl ester of calcein, is highly lipophilic and cell membrane permeable. Although Calcein-AM itself is not a fluorescent molecule, the calcein generated from Calcein- AM by esterase in a viable cell emits a strong green...

Product Description FDA is cell-membrane permeable and accumulates inside of viable cells as fluorescein (Fig. 1). Since fluorescein is less hydrophilic than BCECF or Calcein, the leakage of fluorescein from cells is rather high. FDA is also utilized for flow cytometry. The excitation and emission wavelengths of fluorescein are 488 nm and 530 nm, respectively. FDA-stained cells are shown in Fig. 2. Fig. 1 Cell staining mechanism ...

Product Description Hoechst dyes are cell membrane permeable and stain DNA to emit intense blue fluorescence. They bind to DNA in the minor groove of poly-AT sequence rich areas. Both Hoechst 33342 and Hoechst 33258 are water-soluble and stable in aqueous solutions. The excitation and emission wavelengths of Hoechst-DNA complex are 350 nm and 460 nm, respectively. Chemical Structure ...

Product Description Hoechst dyes are cell membrane permeable and stain DNA to emit intense blue fluorescence. They bind to DNA in the minor groove of poly-AT sequence rich areas. Both Hoechst 33342 and Hoechst 33258 are water-soluble and stable in aqueous solutions. The excitation and emission wavelengths of Hoechst-DNA complex are 350 nm and 460 nm, respectively. Chemical Structure ...

Product Description Propidium iodide (PI) is an ethidium bromide analog that emits red fluorescence upon intercalation with double-stranded DNA. PI does not permeate viable cell membranes, but passes through disturbed cell membranes and stains the nuclei. PI is often used in combination with a fluorescein compound, such as Calcein-AM or FDA, for simultaneous staining of viable and dead cells. The excitation and emission wavelengths of PI-DNA complex are 535 nm and 615 nm, respectively....

Product Description Propidium iodide (PI) is an ethidium bromide analog that emits red fluorescence upon intercalation with double-stranded DNA. PI does not permeate viable cell membranes, but passes through disturbed cell membranes and stains the nuclei. PI is often used in combination with a fluorescein compound, such as Calcein-AM or FDA, for simultaneous staining of viable and dead cells. The excitation and emission wavelengths of PI-DNA complex are 535 nm and 615 nm, respectively....

Product Description The 24-well Plate package* contains 8 plates that are compatible with the TestPiece Holder attached to the Biofilm TestPiece Assay Kit (code: B606). This 24-well Plate package is sold in limited quantities: You can order 1 set of B608 when you order 1 set of Biofilm TestPiece Assay Kit (code: B606). *The 24-well plate is not sold individually. ...

Normally, when the intracellular ATP production decreases, the degradation product ADP will be resynthesized to ATP to maintain the intracellular ATP concentrations. However, when the metabolism of ATP production is disrupted, ATP cannot be resynthesized from ADP, but intracellular ATP is converted to ADP and the ADP/ATP ratio increases. The change in ADP/ATP ratio is not only related to apoptosis and autophagy, but also intracellular energy metabolism, and is used as one of the indicators of...

Amino acids are essential for intracellular protein and nucleic acid synthesis, Especially for cancer cells which are proliferating continually. Since the supply of acetyl CoA from the glycolytic pathway is decreased in cancer cells, they have a furthermore huge demand for amino acids which are an important nutrient source for the TCA cycle. The research has shown that cancer cells increase the expression of the amino acid transporter LAT1 (L-type amino acid transporter 1) and take up a large...

Product Description: Apoptosis is one of the programmed cell death, that plays an important role in maintaining the homeostasis and developmental processes in both plants and animals. Any abnormal cells during the cytogenesis are eliminated by apoptosis. For instance, tumor growth from cancer cells occurred in vivo is inhibited by induction of apoptosis. However, apoptosis is not induced when the error occurs in tumor suppressor gene p53. Thus, the growth of cancer cells has been found to...

Adenosine triphosphate (ATP) is an energy source for living cells and is synthesized by both glycolysis and mitochondrial oxidative phosphorylation. Mitochondria generate 95% of cellular ATP, and mitochondrial dysfunction reduces ATP levels in cells. Therefore, the measurement of ATP levels has been established as a proxy for mitochondrial activity. Decreased ATP levels are associated with cancer, aging, neurodegenerative diseases, and mitochondrial diseases. Cancer cells rely on glycolysis, a...

Product Description BAPTA is a calcium-selective chelator developed by Dr. Tsien. It has logKCa=6.97 and logKMg=1.77. The basic chelating unit resembles that of EGTA, but the two aliphatic nitrogen atoms are replaced by aromatic nitrogen. Thus, BAPTA is not protonated at physiological pH. BAPTA possesses pKa3=5.47 and pKa4=6.36. This property indicates that the deprotonation step is not included in its calcium complexation step, and it has a higher complexation rate than EGTA because it is not...

Product Description BAPTA is a calcium-selective chelator developed by Dr. Tsien. It has logKCa=6.97 and logKMg=1.77. The basic chelating unit resembles that of EGTA, but the two aliphatic nitrogen atoms are replaced by aromatic nitrogen. Thus, BAPTA is not protonated at physiological pH. BAPTA possesses pKa3=5.47 and pKa4=6.36. This property indicates that the deprotonation step is not included in its calcium complexation step, and it has a higher complexation rate than EGTA because it is not...

Product DescriptionBCECF is the most widely used intracellular pH probe. Dr. Tsien and others improved this carboxyfluorescein by introducing two extra carboxylates that allow it to be retained better by the cell. BCECF is highly water-soluble because it has 4 to 5 negative charges at neutral pH; it becomes difficult to pass through the cell membrane after loading. Its pKa value, 6.97, is higher than that of carboxyfluorescein. BCECF has an isosbestic point at 439 nm in the excitation spectra,...

Product Description BCECF is the most widely used intracellular pH probe. Dr. Tsien and others improved this carboxyfluorescein by introducing two extra carboxylates that allow it to be retained better by the cell. BCECF is highly water-soluble because it has 4 to 5 negative charges at neutral pH; it becomes difficult to pass through the cell membrane after loading. Its pKa value, 6.97, is higher than that of carboxyfluorescein. BCECF has an isosbestic point at 439 nm in the excitation...

Description Biofilms are biological aggregates consisting of microorganisms and extracellular polysaccharides that can exist in many environments. The microorganisms in biofilms are highly resistant to antibiotics; therefore, research into medicines and food ingredients that have anti-biofilm activities is a growing area. Our assay kits for the quantitative measurements of biofilm formation/inhibition of biofilm formation (Biofilm Formation Assay Kit) and anti-biofilm drug efficacy (Biofilm...

Product Description Biofilms are found in various environments. They are aggregates composed of microorganisms and their metabolites, such as extracellular polysaccharides. In recent years, there has been an increasing need to evaluate the amount of biofilm formed on materials, as well as an increasing need for drugs that inhibit biofilm formation. The Biofilm TestPiece Assay Kit uses crystal violet staining to measure the difference in biofilm formation between different materials. By using...

Description Biofilms are biological aggregates consisting of microorganisms and extracellular polysaccharides that can exist in many environments. The microorganisms in biofilms are highly resistant to antibiotics; therefore, research into medicines and food ingredients that have anti-biofilm activities is a growing area. Our assay kits for the quantitative measurements of biofilm formation/inhibition of biofilm formation (Biofilm Formation Assay Kit) and anti-biofilm drug efficacy (Biofilm...

Product DescriptionCell Counting Kit-8 (CCK-8) allows sensitive colorimetric assays for the determination of cell viability in cell proliferation and cytotoxicity assays. Dojindo’s highly water-soluble tetrazolium salt, WST-8, is reduced by dehydrogenase activities in cells to give a yellow-color formazan dye, which is soluble in the tissue culture media. The amount of the formazan dye, generated by the activities of dehydrogenases in cells, is directly proportional to the number of living...

Product Description Cell Counting Kit-F(CCK-F) is utilized for the fluorometric determination of living cell numbers. The amount of the fluorescent dye, calcein, hydrolyzed by esterases in cells is directly proportional to the number of viable cells in culture media (Fig. 1). The 96-well microplate CCK-F assay has a detection range of less than 50 cells to more than 25,000 cells per well (Fig. 2). Since esterases and phenol red in the culture medium interfere with the fluorescence measurement,...

Product Description The control system of the cell cycle is closely linked to cell proliferation. Cell division in which one cell produces two daughter cells occurs as part of the cell cycle. The cell cycle can be analyzed by measuring the cells stained for DNA using flow cytometry, followed by analysis of the proportion of stained nuclei in cell cycle phase. The cell cycle is classified into two major phases: interphase and the mitotic (M) phase. Interphase is composed of G1 phase...

Product Description The control system of the cell cycle is closely linked to cell proliferation. Cell division in which one cell produces two daughter cells occurs as part of the cell cycle. The cell cycle can be analyzed by measuring the cells stained for DNA using flow cytometry, followed by analysis of the proportion of stained nuclei in cell cycle phase. The cell cycle is classified into two major phases: interphase and the mitotic (M) phase. Interphase is composed of G1 phase...

Technical Manual Supplemental Manual Product Description Simple Procedure Applicable for Living and Fixed Cells Detection with Flow Cytometry Markers of Senescent Cells Co-staining with DNA Damage marker: Live cells Co-staining with DNA Damage marker: Fixed cells Confocal Quantitative Image Cytometer Recommended Filter Product Description DNA damages of the normal cells are caused by repeated cell division and oxidative stress. Cellular Senescence, a state of irreversible growth arrest, can be...

Product Manual (SG05) Protocol for Combined Analysis (SG05 and C544) Description: This product is a simple detection kit by plate assay for senescence-associated β-galactosidase (SA-β-gal) activity which is used as a marker for senescent cells. By simply adding SPiDER-βGal, a reagent for detection of β-galactosidase, to 96 well plates, this kit allows you to quantify SA-β-gal activity and makes it possible to evaluate multiple specimens.When normalization is done by the results obtained by...

Product Description CFSE is cell-membrane permeable and readily accumulates inside viable cells where it covalently attaches to intracellular proteins (Fig. 1). Hydrolyzed CFSE emits fluorescence and covalently attached fluorescein molecules do not leak from cells. CFSE-labeled cells can be monitored over several weeks in vivo. Therefore, CFSE is utilized for detection of viable cell as well as for the long-term observation of cell activities by fluorescent microscopy. The excitation and...

Cell surface antigens that are specifically expressed in cancer and immune cells have been actively studied for early detection and treatment of cancer. However, many cell surface proteins have low expression levels, and most of them are difficult to detect and analyze by conventional techniques. A fluorescence detection method using a fluorescence-labeled antibody is widely known as a specific detection method for cell surface proteins (fluorescence immunostaining method). However, it is...

Product Description CTC can be used in various ways to detect bacteria, depending on the objective. Detection methods include the agar plate cultivation method, which involves counting colonies that arise from bacteria cultivation; staining bacteria using the fluorescent stain method; testing for bacteria that has potential for growth using the DVC method; and detecting particular bacteria using the FISH method-DNA amplification method. CTC is reduced to CTC formazan (CTF) by electron transfer...

The cystine transporter xCT is a crucial transporter that maintains cellular redox balance by regulating glutathione synthesis via cystine uptake. xCT is highly expressed in several types of cancer cells and is expected to be a therapeutic target for cancer. Therefore, xCT has recently attracted researchers’ attention as one of the targets for cancer treatment. The xCT inhibitors sulfasalazine and erastin are known to reduce intracellular glutathione levels by inhibiting cystine uptake,...

--> Detection Principle Lactate dehydrogenase(LDH) is an enzyme that presents in almost cell types and it catalyzes the oxidation of lactate to pyruvate in the presence of co-enzyme NAD+. Once cells are impaired by stress, injuries, chemicals, or intercellular signals, LDH is rapidly released from the cell membrane. Thus, the measurement of the amount of released LDH from cells is one of the major methods to assess the cell death. Since Dojindo's Cytotoxicity LDH Assay Kit-WST neither reflects...

Product Description DiBAC4(3) is a Bis-oxonol-type, membrane-potential, sensitive dye. According to membrane depolarization detection, when DiBAC distribution in cytosol is increased, the fluorescence intensitiy is increased. Since an argon laser (488 nm) can be used for DiBAC4(3) excitation, it is applicable for flow cytometry and confocal microscopy. Chemical Structure ...

Product Description Endocytosis is a cellular process that involves macromolecules being taken up through a plasma membrane-derived vesicle called an endosome. The endocytic pathway contributes to the maintenance of intracellular homeostasis by bringing in various nutrients to the cell and transporting unwanted components to the lysosome, which acts as a waste disposal system. Recent findings reveal that disruption of endocytosis is related to certain neurodegenerative disorders and immune...

Description ExoIsolator Exosome Isolation Kit can collect exosomes from cell supernatants with a recovery rate equivalent to the ultracentrifugation(UC) method. Science ExoIsolator Exosome Isolation Kit requires only the filtration procedure, unlike the UC, exosomes are obtained quickly without any complicated operations. Feature 1: Easy to use no technique required ExoIsolator Exosome Isolation Kit includes Filter Holder and Isolation Filter can collect exosomes from cell supernatant by...