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Rat Cyt-C(Cytochrome C) ELISA Kit

Cat.No.:E-EL-R0006
Reactivity: Rat

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	0.39-25 ng/mL
Sensitivity	0.23 ng/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Rat Cyt-C in samples. No significant cross-reactivity or interference between Rat Cyt-C and analogues was observed.
Reproducibility	Both intra-CV and inter-CV are < 10%.
Application	This ELISA kit applies to the in vitro quantitative determination of Rat Cyt-C concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Rat Cyt-C. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Rat Cyt-C and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Rat Cyt-C, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Rat Cyt-C. You can calculate the concentration of Rat Cyt-C in the samples by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.

Item	Specifications	Storage
Micro ELISA Plate(Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	-20℃, 6 months
Reference Standard	96T: 2 vials 48T: 1 vial
Concentrated Biotinylated Detection Ab (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL	-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 6 months
Biotinylated Detection Ab Diluent	1 vial, 14 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25×)	1 vial, 30 mL
Substrate Reagent	1 vial, 10 mL	2-8℃(Protect from light)
Stop Solution	1 vial, 10 mL	2-8°C
Plate Sealer	5 pieces
Manual	1 copy
Certificate of Analysis	1 copy

Technical Data

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Rat Cyt-C were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat Cyt-C were tested on 3 different plates, 20 replicates in each plate.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (ng/mL)	1.33	4.00	8.84	1.36	3.69	8.71
Standard deviation	0.08	0.16	0.28	0.09	0.17	0.29
CV (%)	6.02	4.00	3.17	6.62	4.61	3.33

Recovery

The recovery of Rat Cyt-C spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type	Range (%)	Average Recovery (%)
Serum(n=8)	90-105	97
EDTA plasma (n=8)	86-97	91
Cell culture media (n=8)	96-107	102

Linearity

Samples were spiked with high concentrations of Rat Cyt-C and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Synonyms	CytC, CYCS, CYC, HCS, THC4
Research Area	Cancer, Cell Biology, Cardiovascular, Metabolism, Signal transduction

Assay Procedures

	1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C
	2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C
	3. Aspirate and wash the plate for 3 times
	4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	6. Add 50μL Stop Solution
	7. Read the plate at 450nm immediately. Calculation of the results

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Citations

Food & Function (2022) IF: 6.317
Naringenin mitigates thioacetamide-induced hepatic encephalopathy in rats: Targeting JNK/Bax/caspase-8 apoptotic pathway

DOI: 10.1039/D2FO03470K

Sample: liver,brain
Oxidative Medicine and Cellular Longevity (2020) IF: 5.076
Neuroprotective Effect of the Inhibitor Salubrinal after Cardiac Arrest in a Rodent Model

DOI: 10.1155/2020/7468738

PMID: 32064028
Sample: Serum
Frontiers in Pharmacology (2021) IF: 5.811
Trimetazidine Modulates Mitochondrial Redox Status and Disrupted Glutamate Homeostasis in a Rat Model of Epilepsy

DOI: 10.3389/fphar.2021.735165

Sample: brain
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS (2021) IF: 4.013
Mangiferin mitigates di-(2-ethylhexyl) phthalate-induced testicular injury in rats by modulating oxidative stress-mediated signals, inflammatory cascades, apoptotic pathways, and steroidogenesis

DOI: 10.1016/j.abb.2021.108982

PMID: 34400143
Sample: testicular
ASIAN JOURNAL OF ANDROLOGY (2018) IF: 3.259
MicroRNA expression profile in chronic nonbacterial prostatitis revealed by next-generation small RNA sequencing

DOI: 10.4103/aja.aja_97_18

Sample: Serum
BIOLOGICAL TRACE ELEMENT RESEARCH (2021) IF: 3.738
Conivaptan and Boric Acid Treatments in Acute Kidney Injury: Is This Combination Effective and Safe?

DOI: 10.1007/s12011-021-02977-8

PMID: 34676519
Sample: serum
Metabolic Syndrome and Related Disorders (2022) IF: 2.363
Naringenin Prevents Inflammation, Apoptosis, and DNA Damage in Potassium Oxonate-Induced Hyperuricemia in Rat Liver Tissue: Roles of Cytochrome C, NF-κB, Caspase-3, and 8-Hydroxydeoxyguanosine

DOI: 10.1089/met.2022.0028

PMID: 35796694
Sample: Liver tissue
STEROIDS (2022) IF: 2.76
Apoptosis Promoting Activity of Selected Plant Steroid in MRMT-1 Breast Cancer Cell line by Modulating Mitochondrial Permeation Pathway

DOI: 10.1016/j.steroids.2022.109151

Sample: MRMT-1 cell

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