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Glutamate colorimetric assay kit

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The Glutamate Colorimetric Assay Kit is a laboratory tool designed to quantitatively measure the concentration of glutamate in biological samples. The kit provides a colorimetric detection method that allows for the spectrophotometric determination of glutamate levels.

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Lab products found in correlation

Hanks balanced salt solution (hbss), by Thermo Fisher Scientific (1 mentions) Spectramax plus 384 microplate reader, by Molecular Devices (1 mentions) Lenti x concentrator, by Takara Bio (1 mentions) Spectramax i3x multi mode microplate reader, by Molecular Devices (1 mentions) Astrocyte growth medium, by Lonza (1 mentions) Glutamine assay kit, by Abcam (1 mentions) Versamax, by Molecular Devices (1 mentions) Microplate reader, by Tecan (1 mentions) Lactate colorimetric assay kit, by Abcam (1 mentions) Glucose colorimetric assay kit, by Abcam (1 mentions) Glutamine colorimetric assay kit, by Abcam (1 mentions) Aspartate colorimetric assay kit, by Abcam (1 mentions) Nad nadh assay kit, by Merck Group (1 mentions) Imark microplate absorbance reader, by Bio-Rad (1 mentions) Multilabel plate reader, by PerkinElmer (1 mentions) Glutamate assay buffer, by Abcam (1 mentions) Adenosine assay kit, by Abcam (1 mentions) Enliten atp assay system, by Promega (1 mentions) Pierce bca assay, by Thermo Fisher Scientific (1 mentions) Gaba elisa kit, by LSBio (1 mentions)

Spelling variants of this product

Colorimetric assay kit (281 citations) Glutamate Assay Kit (69 citations) Colorimetric assay (66 citations) Glutamate (3 citations) Glutamate dehydrogenase activity colorimetric assay kit (2 citations)

25 protocols using glutamate colorimetric assay kit

1

Astrocyte Glutamate Clearance Capacity Assay

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The glutamate clearance capacity of astrocyte cultures was assessed using the Glutamate Colorimetric Assay Kit (BioVision, USA) according to the manufacturer’s instructions and in comparison to neuronal cultures. Briefly, cells were plated at a density of 105 cells in matrigel-coated 12-well plates one day prior to performing the assay. 24 h after plating, cells were washed once with HBSS (Gibco) and subsequently incubated with HBSS containing 500 μM glutamate. After incubation for 2 h at 37 °C, 20 μl of cell supernatant or medium control were transferred into a 96-well plate, diluted with assay buffer to a volume of 50 μl and incubated with 100 μl of the reaction mix for 30 min at 37 °C. Absorbance of the product was measured at 450 nm using a microplate reader. For quantification of glutamate concentrations, a standard curve was created in each assay based on measurement of cell-free HBSS containing known glutamate concentrations.
Hallmann A.L., Araúzo-Bravo M.J., Mavrommatis L., Ehrlich M., Röpke A., Brockhaus J., Missler M., Sterneckert J., Schöler H.R., Kuhlmann T., Zaehres H, & Hargus G. (2017). Astrocyte pathology in a human neural stem cell model of frontotemporal dementia caused by mutant TAU protein. Scientific Reports, 7, 42991.
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2

Quantification of Glutamate Release by MG-63 Cells

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The release of glutamate into the culture medium by MG-63 cells was measured with a glutamate colorimetric assay kit (BioVision Inc., Milpitas, CA, USA) according to the manufacturer’s procedure. To determine the release of glutamate into the extracellular medium, the cells were plated onto 6-well plates at a density of 105 cells/well, followed by the same protocol of inducing osteoblast differentiation described above. On days 2 and 4, the cultured medium was collected for glutamate assay, and the remaining cells were used for the quantification of proteins. The medium used in the glutamate assay was phenol red-free DMEM.
Yang J.E., Song M.S., Shen Y., Ryu P.D, & Lee S.Y. (2016). The Role of KV7.3 in Regulating Osteoblast Maturation and Mineralization. International Journal of Molecular Sciences, 17(3), 407.
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3

Extracellular Glutamate Quantification

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The Glutamate Colorimetric Assay Kit (BioVision) was used for detection of extracellular glutamate released into the medium. HT-1080 (20,000 cells/well) were seeded overnight and treated with IFNγ (10 ng/mL) for 24 hours. Cells were incubated in serum free DMEM medium containing 5 μM Erastin for 1 hour. 50 μl medium was taken for extracellular glutamate measurement.
Wang W., Green M., Choi J.E., Gijón M., Kennedy P.D., Johnson J.K., Liao P., Lang X., Kryczek I., Sell A., Xia H., Zhou J., Li G., Li J., Li W., Wei S., Vatan L., Zhang H., Szeliga W., Gu W., Liu R., Lawrence T., Lamb C., Tanno Y., Cieslik M., Stone E., Georgiou G., Chan T.A., Chinnaiyan A, & Zou W. (2019). CD8+ T cells regulate tumor ferroptosis during cancer immunotherapy. Nature, 569(7755), 270-274.
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4

Quantifying Intracellular Glutamate Levels

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Intracellular glutamate concentrations were determined using a glutamate colorimetric assay kit, following the manufacturers guidelines (Biovision). Briefly, cells seeded in a 6-well plate were trypsinised, washed three times with ice-cold PBS and resuspended in 200 μL of assay buffer before sonication on ice (Soniprep 150, MSE). Lysates were then centrifuged at 15,000 × g for 10 min at 4 °C and the supernatant taken for analysis. Total intracellular glutamate was normalized to protein concentration.
Greenwood H.E., Edwards R.S., Tyrrell W.E., Barber A.R., Baark F., Tanc M., Khalil E., Falzone A., Ward N.P., DeBlasi J.M., Torrente L., Pearce D.R., Firth G., Smith L.M., Timmermand O.V., Huebner A., George M.E., Swanton C., Hynds R.E., DeNicola G.M, & Witney T.H. (2023). Imaging the master regulator of the antioxidant response in non-small cell lung cancer with positron emission tomography. bioRxiv.
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5

Glutamate Quantification in Cell Lysates

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The glutamate concentration in cell lysates was measured using the Glutamate Colorimetric Assay kit (Biovision) and following manufacturer’s protocol. Briefly, 1 × 106 cells per tested sample were homogenized in 100 μl assay buffer and centrifuged to remove insoluble material. One hundred μl reaction mix was added to the supernatant, standards and background control samples and after 30 min incubation at 37°C, absorbance was measured at 450 nm with a SpectraMax Plus384 MicroPlate Reader (Molecular Devices). The experiment was performed in quadruplicate.
Redis R.S., Vela L.E., Lu W., de Oliveira J.F., Ivan C., Rodriguez-Aguayo C., Adamoski D., Pasculli B., Taguchi A., Chen Y., Fernandez A.F., Valledor L., Van Roosbroeck K., Chang S., Shah M., Kinnebrew G., Han L., Atlasi Y., Cheung L.H., Huang G.Y., Monroig P., Ramirez M.S., Ivkovic T.C., Van L., Ling H., Gafà R., Kapitanovic S., Lanza G., Bankson J.A., Huang P., Lai S.Y., Bast R.C., Rosenblum M.G., Radovich M., Ivan M., Bartholomeusz G., Liang H., Fraga M.F., Widger W.R., Hanash S., Berindan-Neagoe I., Lopez-Berestein G., Ambrosio A.L., Gomes Dias S.M, & Calin G.A. (2016). Allele-specific reprogramming of cancer metabolism by the long non-coding RNA, CCAT2. Molecular cell, 61(4), 520-534.
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6

Astrocyte Glutamate Uptake Assay

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Human primary astrocytes from Lonza (cat. no. CC-2565) were cultured in Lonza Astrocyte Growth Medium (cat. no. CC-3186) according to the manufacturer’s instructions. To force expression of dipeptide repeat proteins in the astrocytes, lentiviruses were produced in HEK 293T cells and concentrated ~80-fold using Lenti-X Concentrator (Clontech). When astrocytes reached 90% confluency, they were transduced with lentiviruses encoding either GFP or GR(50)-GFP. Fluorescence microscopy analysis was used to confirm that 60–80% of astrocytes were transduced in both conditions. To measure glutamate uptake, GFP- or GR(50)-GFP-expressing astrocytes were incubated with 200 mM glutamate in Astrocyte Growth Medium for 2 hours at 37 degrees Celsius in biological quadruplicate. Media samples were collected after 2 hours and glutamate levels were quantified using the BioVision Glutamate Colorimetric Assay Kit (cat. no. K629) according to the manufacturer’s instructions. Colorimetric substrate levels were quantified at 450 nm using a Molecular Devices SpectraMax i3x Multi-Mode Microplate Reader.
Shi Y., Lin S., Staats K.A., Li Y., Chang W.H., Hung S.T., Hendricks E., Linares G.R., Wang Y., Son E.Y., Wen X., Kisler K., Wilkinson B., Menendez L., Sugawara T., Woolwine P., Huang M., Cowan M.J., Ge B., Koutsodendris N., Sandor K.P., Komberg J., Vangoor V.R., Senthilkumar K., Hennes V., Seah C., Nelson A.R., Cheng T.Y., Lee S.J., August P.R., Chen J.A., Wisniewski N., Victor H.S., Belgard T.G., Zhang A., Coba M., Grunseich C., Ward M.E., van den Berg L.H., Pasterkamp R.J., Trotti D., Zlokovic B.V, & Ichida J.K. (2018). Haploinsufficiency Leads to Neurodegeneration in C9ORF72 ALS/FTD Human Induced Motor Neurons. Nature medicine, 24(3), 313-325.
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7

Glutamine Metabolism Pathway Profiling

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Cells were transfected for 48 h and the culture medium was collected. The glutamine consumption was detected using the Glutamine Assay Kit (Biovision, Milpitas, CA, USA) following the manufacturer’s protocols. In addition, alpha-ketoglutarate (α-KG) and glutamate concentrations were respectively measured by Alpha-Ketoglutarate Colorimetric Assay Kit and Glutamate Colorimetric Assay Kit (Biovision).
Chang Z., Fu Y., Jia Y., Gao M., Song L., Zhang W., Zhao R, & Qin Y. (2021). Circ-SFMBT2 drives the malignant phenotypes of esophageal cancer by the miR-107-dependent regulation of SLC1A5. Cancer Cell International, 21, 495.
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8

Glutamate Quantification in Mouse Spinal Cord

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Glutamate concentrations were determined using a Glutamate Colorimetric Assay Kit (K629, BioVision, United States). Glutamate concentrations were measured according to the manufacturer’s instructions. In brief, the lumbar enlargement of the spinal cord was collected from the mice and homogenized in 100 μL of glutamate assay buffer. Then the samples were centrifuged at 13,000 g at 4°C for 10 min, and the supernatants were collected. After the glutamate assay reagents were added and mixed, the reaction was incubated for 30 min at 37°C. A microplate reader (Versa Max, Molecular Devices) was employed to measure the optical density (OD) at 450 nm and glutamate concentrations were calculated.
He L., Xu W., Zhang C., Ding Z., Guo Q., Zou W, & Wang J. (2022). Dysregulation of Vesicular Glutamate Transporter VGluT2 via BDNF/TrkB Pathway Contributes to Morphine Tolerance in Mice. Frontiers in Pharmacology, 13, 861786.
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9

Extracellular Glutamate Quantification

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The Glutamate Colorimetric Assay Kit (BioVision, K629) was used for detection of extracellular glutamate released into the medium. Parental or SLC7A11 knockout B16F10 cells were seeded overnight in the presence of 2-mercaptoethanol to support knockout cell growth. Results normalized to cell viability.
Lang X., Green M.D., Wang W., Yu J., Choi J.E., Jiang L., Liao P., Zhou J., Zhang Q., Dow A., Saripalli A.L., Kryczek I., Wei S., Szeliga W., Vatan L., Stone E.M., Georgiou G., Cieslik M., Wahl D.R., Morgan M.A., Chinnaiyan A.M., Lawrence T.S, & Zou W. (2019). Radiotherapy and immunotherapy promote tumoral lipid oxidation and ferroptosis via synergistic repression of SLC7A11. Cancer discovery, 9(12), 1673-1685.
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10

Extracellular Glutamate Quantification

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The Glutamate Colorimetric Assay Kit (BioVision) was used for detection of extracellular glutamate released into the medium. HT-1080 (20,000 cells/well) were seeded overnight and treated with IFNγ (10 ng/mL) for 24 hours. Cells were incubated in serum free DMEM medium containing 5 μM Erastin for 1 hour. 50 μl medium was taken for extracellular glutamate measurement.
Wang W., Green M., Choi J.E., Gijón M., Kennedy P.D., Johnson J.K., Liao P., Lang X., Kryczek I., Sell A., Xia H., Zhou J., Li G., Li J., Li W., Wei S., Vatan L., Zhang H., Szeliga W., Gu W., Liu R., Lawrence T., Lamb C., Tanno Y., Cieslik M., Stone E., Georgiou G., Chan T.A., Chinnaiyan A, & Zou W. (2019). CD8+ T cells regulate tumor ferroptosis during cancer immunotherapy. Nature, 569(7755), 270-274.
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