Glutamate measurements were performed using the Fluorometric Glutamate Assay Kit from Abcam (ab138883). Samples and assay buffers were prepared in correspondence with manufacturer’s instructions. Briefly, cells were seeded in 6-well plates and treated with indicated doses of CB-839 or vehicle control for 24 hours prior to the experiment. Following treatment, cells were collected and counted for normalization. Cells were lysed in 100 μL of 1× mammalian lysis buffer (Abcam) and incubated for 20 minutes. 50 μL of sample lysate was added to 96-well black-walled plates along with 50 μL of Glutamic acid Reaction Mix prepared according to manufacturers instructions. The plate was incubated at room temperature for 30 minutes and fluorescent intensity of Ex/Em = 540/590 nm was measured using the Synergy 2 microplate reader (BioTek).
Glutamate assay kit fluorometric
Manufactured by Abcam
Sourced in United Kingdom
The Glutamate Assay Kit (Fluorometric) is a laboratory tool designed to quantify glutamate levels in various sample types. It utilizes a fluorometric method to enable sensitive and accurate determination of glutamate concentration.
Automatically generated - may contain errors
Lab products found in correlation
Synergy 2, by Agilent Technologies (1 mentions)
Aspartate assay kit, by Abcam (1 mentions)
L amino acid assay kit, by Abcam (1 mentions)
Aspartate colorimetric fluorometric assay kit, by Abcam (1 mentions)
Atp detection assay kit luminescence, by Cayman Chemical (1 mentions)
Synergy h1 plate reader, by Agilent Technologies (1 mentions)
Victor x4 multilabel plate reader, by PerkinElmer (1 mentions)
6 protocols using glutamate assay kit fluorometric
1
Fluorometric Glutamate Quantification in Cells
2
Measuring Glutamate Release in U87MG Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Parental U87MG (U87MG‐P) or U87MG‐E cells were transferred to 6‐well plates (1 × 106 cells per well), cultured for 12 hours, washed with PBS, and cultured for an additional 8 hours in 2 mL of glutamate‐free medium (DMEM with glucose at 4.5 g/L; Nacalai) in the absence or presence of sulfasalazine (400 μmol/L). The amount of glutamate released into culture supernatants was then measured with the use of a fluorometric glutamate assay kit (Abcam, Tokyo, Japan).
3
Metabolic Analysis of TF-1a Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
TF-1a-Scramble, TF-1a-LIN28B-shRNA3, and TF-1a-LIN28B-shRNA5 cells were lysed in RIPA lysis buffer. Three kits were purchased from Abcam (Cambridge, UK), including Glutamate Assay Kit (Fluorometric) (ab138883), L-Amino Acid Assay Kit (ab65347), and Aspartate Assay Kit (ab102512). The measurement of glutamate, L-amino acid, and aspartate were performed according to manufacturer’s specifications.
4
Inflammatory Cytokine and Glutamate Measurements
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The levels of IFNγ, IL-1β, and IL-6 were measured by M.S.D. 96-well plate Human Pro-Inflammatory V-PLEX Human Pro-Inflammatory Assay kits. The secreted levels of Glutamate were measured by Glutamate Assay Kit (Fluorometric) (Abcam; ab138883).
5
Quantification of Brain Neurotransmitters
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Levels of neurotransmitters including glutamate, aspartate, and GABA in brain tissue were measured.
For the measurement of glutamate levels in brain tissue, brain homogenate samples were incubated with mammalian cell lysis buffer (PerkinElmer) at a 1:1, v/v ratio, and further lysed by shearing mechanical force achieved by passing the mixture through an 18- or 21-gauge needle. The lysates were centrifuged at 2,150 ×g at 4°C for 10 min, and supernatants were analyzed using a Glutamate Assay Kit (Fluorometric) (Abcam).
For the measurement of aspartate levels in brain tissue, brain homogenate samples were combined with 0.1 M HCl at a 1:1 v/v ratio. The mixtures were centrifuged at 2,150 ×g at 4°C for 10 min, and supernatants were analyzed using an aspartate Colorimetric/Fluorometric Assay Kit (BioVision).
For the measurement of GABA levels in the striatum of the brain, striatal tissue was isolated from the whole brain and subjected to homogenization in 0.7 ml of mammalian cell lysis buffer. The samples were further lysed by shearing mechanical force achieved by passage through an 18- or 21-gauge needle. The lysates were centrifuged at 20,000 ×g at room temperature for 5 min, and supernatants were used for the measurement using Mouse GABA ELISA Kit (MyBioSource).
For the measurement of glutamate levels in brain tissue, brain homogenate samples were incubated with mammalian cell lysis buffer (PerkinElmer) at a 1:1, v/v ratio, and further lysed by shearing mechanical force achieved by passing the mixture through an 18- or 21-gauge needle. The lysates were centrifuged at 2,150 ×g at 4°C for 10 min, and supernatants were analyzed using a Glutamate Assay Kit (Fluorometric) (Abcam).
For the measurement of aspartate levels in brain tissue, brain homogenate samples were combined with 0.1 M HCl at a 1:1 v/v ratio. The mixtures were centrifuged at 2,150 ×g at 4°C for 10 min, and supernatants were analyzed using an aspartate Colorimetric/Fluorometric Assay Kit (BioVision).
For the measurement of GABA levels in the striatum of the brain, striatal tissue was isolated from the whole brain and subjected to homogenization in 0.7 ml of mammalian cell lysis buffer. The samples were further lysed by shearing mechanical force achieved by passage through an 18- or 21-gauge needle. The lysates were centrifuged at 20,000 ×g at room temperature for 5 min, and supernatants were used for the measurement using Mouse GABA ELISA Kit (MyBioSource).
6
Astrocyte ATP and Glutamate Kinetics
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For the assessment of ATP and Glu release from astrocytes in the function of time, the cells were seeded onto a 96-well plate to a final density of 1.5 × 104 cells/well for 24 h. Afterwards, the cells were washed and supplied with fresh experimental medium (in case of ATP assessment) or HEPES-buffered solution (HBS; 120 mM NaCl, 5 mM KCl, 1 mM MgCl2, 1.5 mM CaCl2, 10 mM glucose, 25 mM HEPES; pH = 7.4; in case of Glu assessment). After the incubations lasting 2, 5, 10, 15, or 30 min, the supernatants were collected and subjected for the measurands evaluation. For ATP, the luminescence-based test “ATP Detection Assay Kit—Luminescence” (Cayman; Ann Arbor, MI, USA; cat no. 700410) was applied according to the protocol recommended by the producer, and luminescence was assessed with the use of a Synergy H1 plate reader (Biotek; Winooski, VT, USA). For Glu measurement, the fluorescence-based test “Glutamate Assay Kit (Fluorometric)” (Abcam; Cambridge, UK; cat no. ab138883) was used according to the protocol recommended by the producer, and fluorescence (excitation λ = 550 nm, emission λ = 590 nm) was assessed with the use of a Victor X4 multilabel plate reader (PerkinElmer; Waltham, MA, USA).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!