Rat/Mouse Cytochrome c Quantikine ELISA Kit

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MCTC0
Mouse/Rat Cytochrome c ELISA Standard Curve
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Product Details
Procedure
Citations (23)
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Rat/Mouse Cytochrome c Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
2.5 hours
Sample Type & Volume Required Per Well
Cell Lysates (50 uL), Subcellular Fractions (50 uL)
Sensitivity
0.5 ng/mL
Assay Range
0.8 - 25 ng/mL (Cell Lysates, Subcellular Fractions)
Specificity
This kit recognizes both the reduced and oxidized forms of natural rat and mouse cytochrome c.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.

Product Summary

The Quantikine Rat/Mouse Cytochrome c Immunoassay is a 2.5 hour solid phase ELISA designed to measure rat and mouse cytochrome c. It contains natural rat cytochrome c and antibodies raised against cytochrome c, and because of the 100% sequence identity of rat and mouse cytochrome c, this kit can be used to accurately quantitate natural rat or mouse cytochrome c in cell lysates and subcellular fractions.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision

Cell Lysates, Subcellular Fractions

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 1.61 8.82 13.4 1.72 8.91 13.5
Standard Deviation 0.02 0.09 0.19 0.06 0.31 0.59
CV% 1.24 1.02 1.42 3.49 3.48 4.37

Recovery

The recovery of rat/mouse Cytochrome c spiked to three levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
PBS/0.01% Saponin (n=2) 94 90-98
PBS/0.05% Tween 20 (n=2) 101 100-102
PBS/0.5% Triton X-100 (n=2) 100 96-106
PBS/1% Triton X-100 (n=2) 98 95-101

Linearity

To assess the linearity of the assay, five or more samples spiked with various concentrations of rat/mouse Cytochrome c in each matrix were diluted with Calibrator Diluent and then assayed.
Mouse/Rat Cytochrome c ELISA Linearity

Data Examples

Mouse/Rat Cytochrome c ELISA Standard Curve

Product Datasheets

Preparation and Storage

Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Cytochrome c

Cytochrome c is a small, highly conserved, heme-containing protein involved in mitochondrial electron transport. Cytochrome c is released from the mitochondria in response to pro-apoptotic stimuli and serves as a cofactor in initiating the activation of the APAF-1/Caspase-9 complex. The effect of specific proteins and/or small molecules on the initiation of the intrinsic apoptotic pathway can be evaluated in an in vitro system of isolated mitochondria by monitoring the release of cytochrome c.

Entrez Gene IDs:
54205 (Human)
Alternate Names:
CYCHCS; CYCS; Cytochrome c; cytochrome c, somatic; THC4
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Assay Procedure

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 75 µL Conjugate
  4.   Add 75 µL of Conjugate to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

  8. 100 µL Substrate Solution
  9.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

  10. 100 µL Stop Solution
  11.   Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Citations for Rat/Mouse Cytochrome c Quantikine ELISA Kit

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

23 Citations: Showing 1 - 10
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  1. High Concentration of C5a-Induced Mitochondria-Dependent Apoptosis in Murine Kidney Endothelial Cells
    Authors: IJ Tsai, WC Lin, YH Yang, YL Tseng, YH Lin, CH Chou, YK Tsau
    Int J Mol Sci, 2019;20(18):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  2. A Comparison of Mitochondrial Toxicity of Mephedrone on three Separate Parts of Brain including Hippocampus, Cortex and Cerebellum
    Authors: P Naserzadeh, G Taghizadeh, B Atabaki, E Seydi, J Pourahmad
    Neurotoxicology, 2019;0(0):.
    Species: Rat
    Sample Types: Subcellular Fraction
  3. Epigallocatechin-3-gallate attenuates microcystin-LR-induced apoptosis in human umbilical vein endothelial cells through activation of the NRF2/HO-1 pathway
    Authors: J Shi, M Zhang, L Zhang, H Deng
    Environ. Pollut., 2018;239(0):466-472.
    Species: Human
    Sample Types: Cell Lysates
  4. Di-Huang-Yi-Zhi herbal formula attenuates amyloid-?-induced neurotoxicity in PC12 cells
    Authors: HM An, C Lin, C Gu, JJ Chen, WX Sun, M Jin, TL Zhang, MF Qiu, B Hu
    Exp Ther Med, 2017;13(6):3003-3008.
    Species: Rat
    Sample Types: Cell Lysates
  5. microRNA dysregulation in polyglutamine toxicity of TATA-box�binding protein is mediated through STAT1 in mouse neuronal cells
    Authors: R Roshan, A Choudhary, A Bhambri, B Bakshi, T Ghosh, B Pillai
    J Neuroinflammation, 2017;14(1):155.
    Species: Mouse
    Sample Types: Cell Lysates
  6. A Small-Molecule Inhibitor of Bax and Bak Oligomerization Prevents Genotoxic Cell Death and Promotes Neuroprotection
    Authors: X Niu, H Brahmbhatt, P Mergenthal, Z Zhang, J Sang, M Daude, FG Ehlert, WE Diederich, E Wong, W Zhu, J Pogmore, JP Nandy, M Satyanaray, RK Jimmidi, P Arya, B Leber, J Lin, C Culmsee, J Yi, DW Andrews
    Cell Chem Biol, 2017;0(0):.
    Species: Mouse
    Sample Types: Cell Lysates
  7. Allicin Decreases Lipopolysaccharide-Induced Oxidative Stress and Inflammation in Human Umbilical Vein Endothelial Cells through Suppression of Mitochondrial Dysfunction and Activation of Nrf2
    Authors: M Zhang, H Pan, Y Xu, X Wang, Z Qiu, L Jiang
    Cell. Physiol. Biochem., 2017;41(6):2255-2267.
    Species: Human
    Sample Types: Cell Culture Supernates
  8. Epigallocatechin-3-gallate counters cisplatin toxicity of rat testes
    Authors: AA Fouad, HO Qutub, AEA Fouad, AM Audeh, WN Al-Melhim
    Pharm Biol, 2017;55(1):1710-1714.
    Species: Rat
    Sample Types: Tissue Homogenates
  9. Repeated Administration of Mercury Intensifies Brain Damage in Multiple Sclerosis through Mitochondrial Dysfunction
    Authors: F Kahrizi, A Salimi, F Noorbakhsh, M Faizi, F Mehri, P Naserzadeh, N Naderi, J Pourahmad
    Iran J Pharm Res, 2016;15(4):834-841.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  10. Paeoniflorin inhibition of 6-hydroxydopamine-induced apoptosis in PC12 cells via suppressing reactive oxygen species-mediated PKCdelta/NF-kappaB pathway.
    Authors: Dong H, Li R, Yu C, Xu T, Zhang X, Dong M
    Neuroscience, 2015;285(0):70-80.
    Species: Rat
    Sample Types: Cell Lysates
  11. Interleukin-1beta enhances FasL-induced caspase-3/-7 activity without increasing apoptosis in primary mouse hepatocytes.
    Authors: Lutz A, Sanwald J, Thomas M, Feuer R, Sawodny O, Ederer M, Borner C, Humar M, Merfort I
    PLoS ONE, 2014;9(12):e115603.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  12. Dynamin-related protein 1 inhibitors protect against ischemic toxicity through attenuating mitochondrial Ca2+ uptake from endoplasmic reticulum store in PC12 cells.
    Authors: Tian, Ye, Li, Bin, Shi, Wen-Zhen, Chang, Ming-Ze, Zhang, Ge-Juan, Di, Zheng-Li, Liu, Yong
    Int J Mol Sci, 2014;15(2):3172-85.
    Species: Rat
    Sample Types: Cell Culture Supernates
  13. Sirt3 protects cortical neurons against oxidative stress via regulating mitochondrial Ca2+ and mitochondrial biogenesis.
    Authors: Dai, Shu-Hui, Chen, Tao, Wang, Yu-Hai, Zhu, Jie, Luo, Peng, Rao, Wei, Yang, Yue-Fan, Fei, Zhou, Jiang, Xiao-Fan
    Int J Mol Sci, 2014;15(8):14591-609.
    Species: Rat
    Sample Types: Cell Lysates
  14. After embedding in membranes antiapoptotic Bcl-XL protein binds both Bcl-2 homology region 3 and helix 1 of proapoptotic Bax protein to inhibit apoptotic mitochondrial permeabilization.
    Authors: Ding J, Mooers B, Zhang Z, Kale J, Falcone D, McNichol J, Huang B, Zhang X, Xing C, Andrews D, Lin J
    J Biol Chem, 2014;289(17):11873-96.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  15. Iduna protects HT22 cells from hydrogen peroxide-induced oxidative stress through interfering poly(ADP-ribose) polymerase-1-induced cell death (parthanatos).
    Authors: Xu H, Luo P, Zhao Y, Zhao M, Yang Y, Chen T, Huo K, Han H, Fei Z
    Cell Signal, 2013;25(4):1018-26.
    Species: Rat
    Sample Types: Cell Lysates
  16. Inhibition of store-operated calcium entry attenuates MPP(+)-induced oxidative stress via preservation of mitochondrial function in PC12 cells: involvement of Homer1a.
    Authors: Li X, Chen W, Zhang L, Liu W, Fei Z
    PLoS ONE, 2013;8(12):e83638.
    Species: Rat
    Sample Types: Mitochondria
  17. Rapid detection of an ABT-737-sensitive primed for death state in cells using microplate-based respirometry.
    Authors: Clerc, Pascalin, Carey, Gregory, Mehrabian, Zara, Wei, Michael, Hwang, Hyehyun, Girnun, Geoffrey, Chen, Hegang, Martin, Stuart S, Polster, Brian M
    PLoS ONE, 2012;7(8):e42487.
    Species: Mouse
    Sample Types: Cell Lysates
  18. Antimitochondrial autoantibodies in pemphigus vulgaris: a missing link in disease pathophysiology.
    Authors: Marchenko S, Chernyavsky AI, Arredondo J, Gindi V, Grando SA
    J. Biol. Chem., 2010;285(6):3695-704.
    Species: Mouse
    Sample Types: Cell Lysates
  19. Augmenter of liver regeneration: an important intracellular survival factor for hepatocytes.
    Authors: Thirunavukkarasu C, Wang LF, Harvey SA, Watkins SC, Chaillet JR, Prelich J, Starzl TE, Gandhi CR
    J. Hepatol., 2008;48(4):578-88.
    Species: Mouse
    Sample Types: Cell Lysates
  20. Mitochondrially targeted ceramide LCL-30 inhibits colorectal cancer in mice.
    Authors: Dahm F, Bielawska A, Nocito A, Georgiev P, Szulc ZM, Bielawski J, Jochum W, Dindo D, Hannun YA, Clavien PA
    Br. J. Cancer, 2007;98(1):98-105.
    Species: Mouse
    Sample Types: Cell Lysates
  21. Cardiomyocyte-specific Bcl-2 overexpression attenuates ischemia-reperfusion injury, immune response during acute rejection, and graft coronary artery disease.
    Authors: Tanaka M, Nakae S, Terry RD, Mokhtari GK, Gunawan F, Balsam LB, Kaneda H, Kofidis T, Tsao PS, Robbins RC
    Blood, 2004;104(12):3789-96.
    Species: Mouse
    Sample Types: Cell Lysates
  22. Release of intact, monomeric cytochrome c from apoptotic and necrotic cells.
    Authors: Jemmerson R, LaPlante B, Treeful A
    Cell Death Differ., 2002;9(5):538-48.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  23. Posttranslational N-myristoylation of BID as a molecular switch for targeting mitochondria and apoptosis.
    Authors: Zha J, Weiler S, Oh KJ, Wei MC, Korsmeyer SJ
    Science, 2000;290(5497):1761-5.
    Species: Mouse
    Sample Types: Tissue Homogenates

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Rat/Mouse Cytochrome c Quantikine ELISA Kit
By Anonymous on 05/19/2017
Application: Sample Tested: cytochrome c standard