Multiplex Assay Kit for Gamma-Aminobutyric Acid (gABA) ,etc. by CBA (Cytometric Bead Array)

4-Aminobutyric Acid

Specificity

This assay has high sensitivity and excellent specificity for detection of Gamma-Aminobutyric Acid (gABA) ,etc. by CBA (Cytometric Bead Array).
No significant cross-reactivity or interference between Gamma-Aminobutyric Acid (gABA) ,etc. by CBA (Cytometric Bead Array) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of Gamma-Aminobutyric Acid (gABA) ,etc. by CBA (Cytometric Bead Array) and the recovery rates were calculated by comparing the measured value to the expected amount of Gamma-Aminobutyric Acid (gABA) ,etc. by CBA (Cytometric Bead Array) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 84-101 92
EDTA plasma(n=5) 96-105 101
heparin plasma(n=5) 81-95 92

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Gamma-Aminobutyric Acid (gABA) ,etc. by CBA (Cytometric Bead Array) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Gamma-Aminobutyric Acid (gABA) ,etc. by CBA (Cytometric Bead Array) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Gamma-Aminobutyric Acid (gABA) ,etc. by CBA (Cytometric Bead Array) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 92-101% 88-99% 78-101% 78-91%
EDTA plasma(n=5) 79-101% 85-101% 83-104% 91-98%
heparin plasma(n=5) 95-105% 79-98% 79-105% 90-104%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:gABA) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Add 200μL analysis buffer solution to each well of the plate for pre-wetting;
2. Preparation of standards, reagents and samples before the experiment;
3. Add 50μL standard or sample to each well, add 50μL prepared Detection Reagent A, add 10μL magnetic beads, and incubate 90 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
5. Wash plate once on magnetic frame;
6. Add 100μL sheath fluid, swirl for 10 minutes, read on the machine.

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Magazine Citations
Toxicology and Industrial Health Effect of sub-acute exposure to acrylamide on GABAergic neurons and astrocytes in weaning rat cerebellum PubMed: 21444355
Neurosci Lett Expression of the neuron-specific potassium chloride cotransporter KCC2 in adult rat cochlear. PubMed: 18577424
Biomark Insights Decreased Epidermal Growth Factor (EGF) Associated with HMGB1 and Increased Hyperactivity in Children with Autism PubMed: PMC3623607
Biomarker insights Correlation Between Hepatocyte Growth Factor (HGF) and Gamma-Aminobutyric Acid (GABA) Plasma Levels in Autistic Children PubMed: PMC3694825
Biomarker insights Decreased Hepatocyte Growth Factor (HGF) and Gamma Aminobutyric Acid (GABA) in Individuals with Obsessive-Compulsive Disorder (OCD) PubMed: PMC3762604
Scientific Reports Renal Denervation Improves the Baroreflex and GABA System in Chronic Kidney Disease-induced Hypertension pubmed:27917928
Journal of Neuroscience Sex differences in the glutamate signaling pathway in juvenile rats Pubmed:28861894
Scientific Reports A novel homozygous mutation in GAD1 gene described in a schizophrenic patient impairs activity and dimerization of GAD67 enzyme Pubmed: 30341396
Neurochemical Research Aucubin Alleviates Seizures Activity in Li-Pilocarpine-Induced Epileptic Mice: Involvement of Inhibition of Neuroinflammation and Regulation of Neurotransmission Pubmed: 30666488
Neuroscience Letters Differentiation of human glioblastoma U87 cells into cholinergic neuron Pubmed: 30928478
Neurochemical Journal Glutamate Signaling Defects in Propionic Acid Orally Administered to Juvenile Rats as an Experimental Animal Model of Autism
Effect of gamma radiation on combination therapy of certain antiepileptic drugs in rats.
Experimental and Therapeutic Medicine Antiepileptic effects of exogenous β‑hydroxybutyrate on kainic acid‑induced epilepsy Pubmed: 33101467
Exp Anim Dementia model mice exhibited improvements of neuropsychiatric symptoms as well as cognitive dysfunction with neural cell transplantation 33828024
Neurogenetics Age-dependent neurological phenotypes in a mouse model of PRRT2-related diseases 34101060
Sci Rep Overexpression of wild-type human amyloid precursor protein alters GABAergic transmission 34475508
J Food Biochem Adaptogenic potential of ginsenosides against domoic acid‐induced toxicity by regulating neuronal stress and kinate receptors: Ex vivo and in silico studies Pubmed:35102569
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