ELISA Kit for Glial Cell Line Derived Neurotrophic Factor (GDNF) Rattus norvegicus (Rat) Sandwich ELISA

ATF1; ATF2; HFB1-GDNF; Astrocyte-Derived Trophic Factor; Glial Derived Neurotrophic Factor

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  • ELISA Kit for Glial Cell Line Derived Neurotrophic Factor (GDNF) Packages (Simulation)
  • ELISA Kit for Glial Cell Line Derived Neurotrophic Factor (GDNF) Packages (Simulation)
  • ELISA Kit for Glial Cell Line Derived Neurotrophic Factor (GDNF) Results demonstration
  • SEA043Ra.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Glial Cell Line Derived Neurotrophic Factor (GDNF) and the recovery rates were calculated by comparing the measured value to the expected amount of Glial Cell Line Derived Neurotrophic Factor (GDNF) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 95-105 101
EDTA plasma(n=5) 96-103 101
heparin plasma(n=5) 93-105 99

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Glial Cell Line Derived Neurotrophic Factor (GDNF) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Glial Cell Line Derived Neurotrophic Factor (GDNF) 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 Glial Cell Line Derived Neurotrophic Factor (GDNF) 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) 91-101% 90-98% 82-95% 93-101%
EDTA plasma(n=5) 78-103% 79-94% 79-95% 94-103%
heparin plasma(n=5) 81-90% 89-103% 99-105% 92-105%

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 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

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

ELISA Kit for Glial Cell Line Derived Neurotrophic Factor (GDNF)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Glial Cell Line Derived Neurotrophic Factor (GDNF). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Glial Cell Line Derived Neurotrophic Factor (GDNF). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Glial Cell Line Derived Neurotrophic Factor (GDNF), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Glial Cell Line Derived Neurotrophic Factor (GDNF) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Efficient Generation of Functionally Active Spinal Cord Neurons from Spermatogonial Stem Cellspubmed:27566610
  • Safflower Extract and Aceglutamide Injection Promoting Recovery of Peripheral Innervations via Vascular Endothelial Growth Factor-B Signaling in Diabetic MicePMC5717862
  • Acupuncture Alters Pro-infl ammatory Cytokines in the Plasma of Maternally Separated Rat Pupspubmed:28986807
  • GDNF serum levels are found to be lower in opioid-maintained patientsISSN 2531-4122
  • Effects of 220 MHz Pulsed Modulated Radiofrequency Field on the Sperm Quality in RatsPubmed: 30974849
  • The Effects of Nordic Walking With Poles With an Integrated Resistance Shock Absorber on Cognitive Abilities and Cardiopulmonary Efficiency in …Pubmed: 33192480
  • Increased serum brain-derived neurotrophic factor and adrenocorticotropic hormone levels are associated with obsessive compulsive disorder in medication?free?¡­33389158
  • The effect of biodegradable silk fibroin-based scaffolds containing glial cell line-derived neurotrophic factor (GDNF) on the corneal regeneration process34119551
  • Silymarin constrains diacetyl-prompted oxidative stress and neuroinflammation in rats: involvements of Dyn/GDNF and MAPK signaling pathwayPubmed:35366745
  • Umbilical cord-derived mesenchymal stem cell conditioned medium reverses neuronal oxidative injury by inhibition of TRPM2 activation and the JNK signaling …Pubmed:35585377

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