Rat/Mouse Cytochrome c Quantikine ELISA Kit

(16 citations)
(1 Review)
  
  • 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
Sample123123
n202020202020
Mean1.618.8213.41.728.9113.5
Standard Deviation0.020.090.190.060.310.59
CV%1.241.021.423.493.484.37

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.
Rat/Mouse Cytochrome c Quantikine ELISA Kit
Preparation and Storage
  • 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;
    Related Research Areas
    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:

    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.

    16 Citations: Showing 1 - 10
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    Species
    Sample Type
    1. 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 Type: Cell Lysates
    2. 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 Type: Cell Culture Supernates
    3. 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 Type: Cell Lysates
    4. 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 Type: Cell Culture Supernates
    5. 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 Type: Cell Culture Supernates
    6. 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 Type: Cell Culture Supernates
    7. 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 Type: Cell Lysates
    8. 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 Type: Mitochondria
    9. 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 Type: Cell Lysates
    10. 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 Type: Cell Lysates
    11. 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 Type: Cell Lysates
    12. 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 Type: Cell Lysates
    13. 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 Type: Cell Lysates
    14. 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 Type: Cell Lysates
    15. 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 Type: Cell Culture Supernates
    16. 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 Type: Tissue Homogenates
    Expand to show all 16 Citations
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    Summary

    Sample Testedcytochrome c standard

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