ELISA Kit for Cytochrome C (CYCS) Rattus norvegicus (Rat) Sandwich ELISA

Cyt-C; HCS; CYC; CytC

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  • ELISA Kit for Cytochrome C (CYCS) Packages (Simulation)
  • ELISA Kit for Cytochrome C (CYCS) Packages (Simulation)
  • ELISA Kit for Cytochrome C (CYCS) Results demonstration
  • SEA594Ra.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 87-97 92
EDTA plasma(n=5) 93-102 97
heparin plasma(n=5) 93-102 97

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Cytochrome C (CYCS) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Cytochrome C (CYCS) 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 Cytochrome C (CYCS) 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) 81-94% 88-97% 82-101% 97-105%
EDTA plasma(n=5) 94-102% 86-104% 98-105% 84-98%
heparin plasma(n=5) 80-101% 80-105% 89-98% 96-105%

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 Cytochrome C (CYCS)

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 Cytochrome C (CYCS). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Cytochrome C (CYCS). 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 Cytochrome C (CYCS), 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 Cytochrome C (CYCS) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Danhong injection protects cardiomyocytes against hypoxia/reoxygenation- and H2O2-induced injury by inhibiting mitochondrial permeability transition pore openingPubMed: 26320687
  • Gypenosides alleviate myocardial ischemia-reperfusion injury via attenuation of oxidative stress and preservation of mitochondrial function in rat heartPubmed:26800973
  • Clematichinenoside (AR) Attenuates Hypoxia/Reoxygenation-Induced H9c2 Cardiomyocyte Apoptosis via a Mitochondria-Mediated Signaling PathwayPubmed:27248986
  • A polysaccharide (PNPA) from Pleurotus nebrodensis ameliorates hepatic ischemic/reperfusion (I/R) injury in ratspubmed:28709897
  • Plasma Cellular Hypoxia, Mitochondrial Dysfunction, Disease Risk and Prognostic Factors in Type 2 Diabetic Patients in Saudi Arabia.
  • Taxifolin protects rat against myocardial ischemia/reperfusion injury by modulating the mitochondrial apoptosis pathwayPubmed: 30723634
  • High Concentrations of Boric Acid Trigger Concentration-Dependent Oxidative Stress, Apoptotic Pathways and Morphological Alterations in DU-145 Human Prostate …Pubmed: 31066018
  • The Role of Oxidative Stress, Renal Inflammation, and Apoptosis in Post Ischemic Reperfusion Injury of Kidney Tissue: the Protective Effect of Dose-Dependent Boric …
  • Betaine suppresses cell proliferation by increasing oxidative stress–mediated apoptosis and inflammation in DU-145 human prostate cancer cell linePubmed: 31368044
  • Concentration-Dependent Effects of Zinc Sulfate on DU-145 Human Prostate Cancer Cell Line: Oxidative, Apoptotic, Inflammatory, and Morphological AnalyzesPubmed: 31463762
  • Curcumin and LOXblock-1 ameliorate ischemia-reperfusion induced inflammation and acute kidney injury by suppressing the semaphorin-plexin pathway.Pubmed: 32603817
  • Concanavalin A induces apoptosis in a dose©\dependent manner by modulating thiol/disulfide homeostasis in C6 glioblastoma cells33604990
  • Reproductive Effects of Nicotinamide on Testicular Function and Structure in Old Male Rats: Oxidative, Apoptotic, Hormonal, and Morphological Analyses34101148
  • Comparative effects of metformin and Cistus laurifolius L. extract in streptozotocin-induced diabetic rat model: oxidative, inflammatory, apoptotic, and histopathological?¡­34097215
  • Allicin ameliorates renal ischemia/reperfusion injury via inhibition of oxidative stress and inflammation in rats34426252
  • 12/15–Lipoxygenase debilitates mitochondrial health in intermittent hypobaric hypoxia induced neuronal damage: An in vivo studyPubmed:34979449
  • In Vivo and In Vitro Enhanced Tumoricidal Effects of Metformin, Active Vitamin D3, and 5-Fluorouracil Triple Therapy against Colon Cancer by Modulating the …Pubmed:35326689
  • Proteomic analysis reveals USP7 as a novel regulator of palmitic acid-induced hepatocellular carcinoma cell deathPubmed:35732625
  • Microbial hydrogen “manufactory” for enhanced gas therapy and self-activated immunotherapy via reduced immune escapePubmed:35705974
  • Oksidatif, Apoptotik ve İnflamatuar Sinyal Yolakları üzerinden C6 Glioblastoma Hücrelerindeki ML351'in Antiproliferatif Etkileri
  • Metformin and Calcitriol Enhance 5-Fluorouracil Tumoricidal Effects in Colon Cancer By Modulating The PI3K/Akt/PTEN/mTOR Network

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