Horse cytochrome c

Manufactured by Merck Group

Sourced in United States

Horse cytochrome c is a laboratory reagent used in biochemical and biophysical research. It is a small heme-containing protein that functions as an electron transport component in the mitochondrial respiratory chain. The core function of horse cytochrome c is to facilitate the transfer of electrons between complex III and complex IV in the electron transport system.

Automatically generated - may contain errors

Lab products found in correlation

Cd11c microbeads, by Miltenyi Biotec (2 mentions) Horse myoglobin, by Merck Group (2 mentions) Dq ovalbumin dq ova, by Thermo Fisher Scientific (1 mentions) Pam3cysserlys4 pam3csk4, by InvivoGen (1 mentions) Alexa fluor 647 af647, by Thermo Fisher Scientific (1 mentions) Rmil 4, by R&D Systems (1 mentions) Polyinosinic polycytidylic acid poly 1 c, by InvivoGen (1 mentions) Decylubiquinone, by Merck Group (1 mentions) Multiscan sky, by Thermo Fisher Scientific (1 mentions) Cytochrome c, by Merck Group (1 mentions) Gelfree system, by Abcam (1 mentions) Bovine ubiquitin, by Merck Group (1 mentions) Ethylenediamine tetraacetic acid edta, by Beijing Chemical Works (1 mentions) 5 furd, by Merck Group (1 mentions) Thioflavin t, by Merck Group (1 mentions) Naringin, by Merck Group (1 mentions) Superdex 75 10 300 column, by GE Healthcare (1 mentions) Kta system, by Cytiva (1 mentions) Human hemoglobin, by Merck Group (1 mentions)

9 protocols using horse cytochrome c

Immunophenotyping and Cellular Assays

Check if the same lab product or an alternative is used in the 5 most similar protocols

Fluorescently labeled antibodies including: Phycoerythrin (PE)-labeled α-CXCR4 (2B11), Brilliant violet 421-labeled α-CD8 (53-6.7), allophycocyanin (APC)-labeled α-CD3ε (BM8), and APC-labeled α-T-bet (4B10), PE-labeled α-granzyme B (NGZB), PE-labeled α-interferon gamma (IFNγ) (XMG1.2), APC labeled α-TNFα (MP6-XT22) [eBioscience (San Diego, CA)] as well as PE-labeled α-H2K^d [Biolegend (San Diego, CA)] were used. Recombinant mouse granulocyte-macrophage colony stimulating factor (rmGM-CSF) and rmIL-4 were purchased from R&D systems (Minneapolis, MN). Pam3CysSerLys4 (Pam3CSK4) and Polyinosinic-polycytidylic acid (Poly(I:C)) were purchased from Invivogen (San Diego, CA). Lipopolysachharide (LPS) was purchased from Sigma (St. Louis, MO). Polybead amino 3.0 μm microspheres were purchased from Polysciences (Warrington, PA). Horse cytochrome c was purchased from Sigma. Alexa Fluor 647 (AF647) and DQ ovalbumin (DQ-OVA) were purchased from Life technologies (Grand Island, NY). Fluorescein isothiocyanate (FITC) conjugated ovalbumin (OVA) was purchase from Sigma.

Liu C., Whitener R.L., Lin A., Xu Y., Chen J., Savinov A., Leiding J.W., Wallet M.A, & Mathews C.E. (2019). Neutrophil Cytosolic Factor 1 in Dendritic Cells Promotes Autoreactive CD8+ T Cell Activation via Cross-Presentation in Type 1 Diabetes. Frontiers in Immunology, 10, 952.

+ Open protocol

+ Expand

Purification and Activity Assay of Arabidopsis I+III2 Supercomplex

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Arabidopsis I + III₂ supercomplex purified by sucrose gradient ultracentrifugation was tested for activity, using an established protocol^{59 (link)}. The assay was carried out in 25 mM potassium phosphate (pH 7.2), 5 mM magnesium chloride, 260 µM NADH, 67 µM decylubiquinone (Sigma-Aldrich), 0.1 mM horse cytochrome c (Sigma-Aldrich) and 3,000 U ml⁻¹ superoxide dismutase to ensure that cytochrome c remained oxidized^{25 (link)}. The assay volume was 170 µl. Four different conditions with or without cytochrome c and decylubiquinone were tested (Supplementary Fig. 5). For each condition, measurements of two blanks (0 µg I + III₂ supercomplex) as well as two measurements each of 2 µg and 4 µg I + III₂ supercomplex were performed. Activity measurements were carried out with a plate reader (Multiscan Sky, Thermo Fisher Scientific) at 340 nm (NADH absorbance; extinction coefficient of NADH: 6.22 mM⁻¹ cm⁻¹).

Klusch N., Dreimann M., Senkler J., Rugen N., Kühlbrandt W, & Braun H.P. (2022). Cryo-EM structure of the respiratory I + III2 supercomplex from Arabidopsis thaliana at 2 Å resolution. Nature Plants, 9(1), 142-156.

+ Open protocol

+ Expand

Enrichment and Stimulation of CD11c+ Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Pulmonary CD11c⁺ cells were enriched by positive selection using CD11c⁺MicroBeads (Miltenyi Biotec). 100,000 CD11c⁺ cells per well were plated in 96-well plates overnight in RPMI-10% FCS with or without 10 mg per ml horse cytochrome c (Sigma), along with PBS, 10 μg per ml Poly I:C or 50 μg per ml R848.

Desch A.N., Gibbings S.L., Clambey E.T., Janssen W.J., Slansky J.E., Kedl R.M., Henson P.M, & Jakubzick C. (2014). Dendritic cell subsets require cis-activation for cytotoxic CD8 T cell induction. Nature communications, 5, 4674.

+ Open protocol

+ Expand

Protein Fractionation Using Sephadex G-75

Check if the same lab product or an alternative is used in the 5 most similar protocols

Blood serum samples (2 ml) or cytosolic fractions (2 ml containing about 40 mg total protein) were fractionated on the column with Sephadex G-75 in phosphate saline buffer, pH 7.4 or buffer A, respectively. The void volume of the column was estimated using blue dextran. Horse cytochrome C was used as a molecular weight marker (Sigma, USA). The fractions following the void volume (~1.5 ml per fraction) were collected and specified by D₂₈₀ and D₂₅₄.

Zatulovskaia Y.A., Ilyechova E.Y, & Puchkova L.V. (2015). The Features of Copper Metabolism in the Rat Liver during Development. PLoS ONE, 10(10), e0140797.

+ Open protocol

+ Expand

Enrichment and Stimulation of CD11c+ Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

+ Open protocol

+ Expand

Yeast Proteome Fractionation and Analysis

Cited 2 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

Bovine Ubiquitin (UniProt Accession P0CG53), horse Cytochrome C (UniProt Accession L7MRG1), and horse myoglobin (UniProt Accession P68082) were purchased as standards from Sigma. Each sample was dissolved to yield solutions of ~10 pmol/μL in 49.9:49.9:0.2 acetonitrile/water/formic acid for infusion studies.
Yeast cells (Saccharomyces cerevisiae strain Y1788) were grown to log phase (OD₆₀₀ = 0.7) at which time they were washed, pelleted, snap-frozen in liquid nitrogen, and stored at −80 °C until use. Yeast cells were lysed by heat and proteins were precipitated in acetone. The proteins were then dissolved in 1% sodium dodecyl sulfate (SDS) and separated by molecular weight (MW) using a GELFrEE system (Expedeon).^{7 (link),26 (link)} Approximately 400 μg of protein were collected into 11 fractions. Two of the fractions were selected for top-down analyses. Prior to mass spectrometric analyses, sodium dodecyl sulfate was removed from the fractions via methanol–chloroform precipitation and proteins were reconstituted with 5% acetonitrile (ACN) and 0.2% formic acid in water.

Lu L., Scalf M., Shortreed M.R, & Smith L.M. (2021). Mesh fragmentation improves dissociation efficiency in top-down proteomics. Journal of the American Society for Mass Spectrometry, 32(6), 1319-1325.

+ Open protocol

+ Expand

Bovine Cu2Zn2SOD1 Extraction and Isoflavone Interactions

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cu₂Zn₂SOD1 from bovine erythrocytes was purchased from Beyotime Biotechnology (Shanghai, China). Daidzein, genistein, sophricoside, genistin, and (purity ≥98% for all) were provided from Nature Institutes for Food and Drug Control(Beijing, China). Glycitein, daidzin, and flycitin were purchased from Chengdu Herbpurify Co., LTD (Chengdu, China). Horse cytochrome c, horse myoglobin, naringin, Thioflavin-T (ThT), ammonium acetate, and 5-furd were obtained from Sigma-Aldrich (St. Louis, MO, USA). Trifluoroethanol (TFE) was purchased from J&K Scientific Ltd. (Beijing, China). Ethylenediaminetetraacetic acid (EDTA) and copric chloride dihydrate (CuCl₂.2H₂O) were obtained from Beijing Chemical Works (Beijing, China). Methanol and formic acid were supplied by TEDIA Company (Fairfield, OH, USA). The ultrapure water used in all experiments was prepared by a Milli-Q water purification system (Milford, MA, USA). The dialysis devices (Micro Float-A-Lyzer, MW-cut: 10 kDa) were bought from Spectrum Laboratories (Rancho Dominguez, CA, USA). The isoflavones, 5-furd, and naringin were dissolved at 1 mM in methanol as stock solutions and stored at 4 °C before experiments.

Bian X., Zhuang X., Xing J., Liu S., Liu Z, & Song F. (2022). Native Mass Spectrometry Coupled to Spectroscopic Methods to Investigate the Effect of Soybean Isoflavones on Structural Stability and Aggregation of Zinc Deficient and Metal-Free Superoxide Dismutase. Molecules, 27(21), 7303.

+ Open protocol

+ Expand

Size-Exclusion Chromatography for Protein Quaternary Structure

Check if the same lab product or an alternative is used in the 5 most similar protocols

Quaternary structure of BpsγCA was investigated by
SEC as previously described [49] (link). In particular, column
calibration curve was performed on a Superdex 75 10/300 column (GE
Healthcare) connected to an ÄKTA™ System (Cytiva) at room temperature.
Running buffer was prepared with 20 mM Tris, 100 mM NaCl, pH 8.0.
Calibration was carried out using the following standards (Sigma Aldrich,
St. Louis, MO, USA): horse Cytochrome C (Cyt C,
12.4 kDa), Bovine Serum Albumin (BSA, 66.4 kDa) and Carbonic Anhydrase from
bovine erythrocytes (CA 29.0 kDa). Blue dextran (2,000,000 Da) was used to
calculate the column void volume (Vo). The molecular weight of BpsγCA was
determined by plotting K_av, calculated from the measured
elution volume (Eq. (3)) against the logarithm of the molecular weights of
the standard proteins. Prism - GraphPad software was used to generate the
graph [50] .

K_{av} = (V_{e} - V_{0}) / (V_{t} - V_{0})

Di Fiore A., De Luca V., Langella E., Nocentini A., Buonanno M., Monti S.M., Supuran C.T., Capasso C, & De Simone G. (2022). Biochemical, structural, and computational studies of a γ-carbonic anhydrase from the pathogenic bacterium Burkholderia pseudomallei. Computational and Structural Biotechnology Journal, 20, 4185-4194.

+ Open protocol

+ Expand

Purification and Polyclonal Antibody Generation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human hemoglobin, cytoglobin and myoglobin and horse cytochrome C were obtained from a commercial source (Sigma-Aldrich). Polyclonal rabbit antisera for detection of PqqL, SurA, BamA and YtfP were generated at the Monash Animal Research Platform, from proteins purified in-house. Rabbits were serially injected with purified proteins in combination with complete (first injection) or incomplete (subsequent injections) Freund's adjuvant, over a period of 1–3 months, with rabbit serum periodically tested for reactivity to the target protein. Once acceptable levels of reactivity were achieved rabbits were sacrificed and serum was collected and stored in aliquots at -80°C.

Grinter R., Leung P.M., Wijeyewickrema L.C., Littler D., Beckham S., Pike R.N., Walker D., Greening C, & Lithgow T. (2019). Protease-associated import systems are widespread in Gram-negative bacteria. PLoS Genetics, 15(10), e1008435.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!