ELISA Kit for Glutathione (GSH)

L-Reduced Glutathione

Specificity

This assay has high sensitivity and excellent specificity for detection of Glutathione (GSH).
No significant cross-reactivity or interference between Glutathione (GSH) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of Glutathione (GSH) and the recovery rates were calculated by comparing the measured value to the expected amount of Glutathione (GSH) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 83-105 87
EDTA plasma(n=5) 86-93 89
heparin plasma(n=5) 88-104 97

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Glutathione (GSH) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Glutathione (GSH) 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 Glutathione (GSH) 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) 94-103% 90-102% 79-104% 97-105%
EDTA plasma(n=5) 94-103% 94-102% 97-104% 82-92%
heparin plasma(n=5) 94-101% 78-101% 90-97% 81-96%

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
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1 Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
Reagent Diluent 1×300µL Stop Solution 1×6mL
TMB Substrate 1×9mL Instruction manual 1
Wash Buffer (30 × concentrate) 1×20mL

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

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Magazine Citations
European Journal of Clinical Nutrition (2012) 66 Evidence for augmented oxidative stress in the subjects with type 1 diabetes and their siblings: a possible preventive role for antioxidants PubMed: 22781023
International Journal of Medical Sciences Glutathione S-Transferase P1 Correlated with Oxidative Stress in Hepatocellular Carcinoma PubMed: PMC3619117
Reproduction Alteration in the intrafollicular thiol–redox system in infertile women with endometriosis Pubmed:25376627
Iran J Basic Med Sci Effects of mild hypothermia therapy on the levels of glutathione in rabbit blood and cerebrospinal fluid after cardiopulmonary resuscitation PubMed: 25810895
Bulletin of the Veterinary Institute in Pulawy Oxidative Stability of Ostrich Meat Related to Duration of Linseed and Lucerne Supplementation to the Bird&#039;s Diet View: J
Journal of Allergy and Clinical Immunology Prenatal maternal distress affects atopic dermatitis in offspring mediated by oxidative stress Pubmed:27016803
Oxidative Medicine and Cellular Longevity Hydrogen sulfide protects against chronic unpredictable mild stress-induced oxidative stress in hippocampus by upreCavia (Guinea pig )lation of BDNF-TrkB pathway journals:2153745.pdf
Institutional repository of Vilnius University Rapid opioid detoxification with an implementation of the new method of gradually increasing dosage of naltrexone induction object:elaba:15733325
Molecular Medicine Reports Toxicity study of oxalicumone A, derived from a marine-derived fungus Penicillium oxalicum, in cultured renal epithelial cells pubmed:28260084
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