COXII | Plant Cytochrome oxidase subunit II (affinity purified)

Product no: AS04 053A

AS04 053A | Clonality: Polyclonal  |  Host: Rabbit  |  Reactivity: Eudicots, monocots, Physcomitrella patens  |  Cellular [compartment marker] of mitochondrial inner membrane

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  • Product Info
  • Immunogen:

    KLH-conugated synthetic peptide fully conserved in all available protein sequences from eudicots including Arabidopsis thaliana AtmG00160, monocots including  Oryza sativa P04373 and Physcomitrella patens Q1XGA9

    Host: Rabbit
    Clonality: Polyclonal
    Purity: Immunogen affinity purified serum in PBS pH 7.4.
    Format: Lyophilized
    Quantity: 50 µg
    Reconstitution: For reconstitution add 50 µl of sterile water
    Storage: Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
    Tested applications: Blue Native-PAGE (BN-PAGE), Western blot (WB)
    Recommended dilution: 1 : 1000 (BN-PAGE), 1 : 1000 (WB)
    Expected | apparent MW:

    29.4 | 30 kDa (for Arabidopsis thaliana)

  • Reactivity
  • Confirmed reactivity: Actinidia chinensis, Arabidopsis thaliana (leaf extract and isolated mitochondria), Betula nana, Brassica napus, Brassica oleracea, Cicer aretinum, Cucumis melo, Cucumis sativus, Erophorum vaginatum, Hordeum vulgare, Lilium longifolorum, Nicotiana tabacum, Picea abies, Plantago major, Plantago euryphylla, Silene uniflora, Silene dioica, Physcomitrium patens, Triticum aestivum, Triticum durum Desf. , Zea mays, Vicia faba, Quercus rubra
    Predicted reactivity: Cucumis melo, Glycine max, Gossypium hirsutum, Oryza sativa, Physcomitrium patens, Pisum sativum, Triticum aestivum, Vigna radiata
    Species of your interest not listed? Contact us
    Not reactive in: Saccharina japonica
  • Application Examples
  • western blot detection using anti-COXII antibody

    1.8 µg of total protein from (1) Arabidopsis thaliana leaf , (2) Plantago major leaf, (3) Plantago europhylla leaf, (4) Silene dioica leaf, (5) Silene uniflora leaf were separated on  4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to PVDF. Primary antibody was used in 1: 1000 dilution. Detection was performed using chemiluminescence, following manufacture's recommendations.

    Application examples:

    Reactant: Oryza sativa (Asian rice)

    Application: Western Blotting

    Pudmed ID: 28056797

    Journal: BMC Plant Biol

    Figure Number: 2A

    Published Date: 2017-01-05

    First Author: Yang, N. & Wang, T.

    Impact Factor: 4.142

    Open Publication

    Plasma membrane (PM) purity verification and PM-related proteins evaluation. a Western blot examination of plasma membrane enrichment. Proteins from entire cell (EC) lysates, microsomal vesicles (MSV), PM (plasma membrane vesicles) and carbonate-washed PM (CPM) were separated by 10% SDS-PAGE, transferred to PVDF membranes and detected with antibodies for PMA2, a PM marker; COXII, a mitochondrial marker; V-ATPase, a vacuole marker; S14-1, a ribosome marker; or H1, a nucleus marker. For detection of PMA2, COXII and V-ATPase, 5 ?g protein was loaded per lane; for detection of S14-1 and H1, 10 ?g protein was loaded per lane. b Summary of proteins with transmembrane domain (TMD) predicted with use of HMMTOP 2.0. c Venn diagram depicting the distribution of proteins with different lipid modifications. GPI, glycosylphosphatidylinositol anchor; Pre, prenylation site; Myr, myristoylation site; Pal, palmitoylation site. d Proteins predicted to have a TMD or post-translational modification (PTM) or both. e Protein subcellular locations annotated by Gene Ontology or WoLF PSORT showed that 1,121 of the 1,797 proteins with a TMD and/or PTM had PM location information


    Reactant: Cucumis sativus (Cucumber)

    Application: Western Blotting

    Pudmed ID: 30304441

    Journal: J Exp Bot

    Figure Number: 2B

    Published Date: 2019-01-01

    First Author: Migocka, M., Maciaszczyk-Dziubinska, E., et al.

    Impact Factor: 6.088

    Open Publication

    Subcellular localization of CsMTP6. (A) Mitochondrial localization of CsMTP6 in protoplasts prepared from suspensions of Arabidopsis cells: 1, GFP fluorescence of protoplast expressing CsMTP6-GFP; 2, MitoTracker red fluorescence; 3, merged image; 4, transmission image of the same protoplast. The scale bar is 10 µm. (B) Immunolocalization of CsMTP6 in cucumber root cells. SDS-PAGE followed by western blot analysis of plasma membrane (PM), tonoplast (Ton), and mitochondrial (Mit) and plastidial (Plast) fractions. The fractions (15 ?g of protein per lane) were blotted with the primary antibodies to identify marker enzymes for the plasma membrane (PM-ATPase), tonoplast (V-ATPase), plastids (Toc75), mitochondria (COXII), and to localize CsMTP6. L, protein ladder.


    Reactant: Cucumis sativus (Cucumber)

    Application: Western Blotting

    Pudmed ID: 30304441

    Journal: J Exp Bot

    Figure Number: 7A

    Published Date: 2019-01-01

    First Author: Migocka, M., Maciaszczyk-Dziubinska, E., et al.

    Impact Factor: 6.088

    Open Publication

    Effects of CsMTP6 expression on mitochondrial Fe and Mn contents and Mn-SOD activity in Arabidopsis protoplasts. (A) Western blot analysis of the purity of mitochondrial and plastidial fractions prepared from protoplasts expressing CsMTP6 using primary antibodies raised against GFP (to detect CsMTP6-GFP), and marker proteins for plastids (Toc75) and mitochondria. The mitochondria and plastid fractions were prepared from protoplasts by discontinuous density gradient centrifugation in Percoll. (B) Fe and (C) Mn content in the mitochondrial fraction prepared from Arabidopsis protoplasts expressing CsMTP6 or transformed with the empty vector. Data are means (±SD) from three separate experiments. (D) Mitochondrial Mn-SOD activity in protoplasts expressing CsMTP6. Data are means (±SD) from three separate experiments, expressed as enzyme units (U). Significant differences were determined using Student’s t-test.


    Reactant: Cucumis sativus (Cucumber)

    Application: Western Blotting

    Pudmed ID: 30304441

    Journal: J Exp Bot

    Figure Number: 8B

    Published Date: 2019-01-01

    First Author: Migocka, M., Maciaszczyk-Dziubinska, E., et al.

    Impact Factor: 6.088

    Open Publication

    The levels of CsMTP6 transcript and protein under different Fe and Mn availability. (A) Real-time analysis of CsMTP6 expression in the roots of 2-week-old cucumber seedlings growing under control Fe (75 µM), control Mn (0.1 µM), excess Fe (1 mM), excess Mn (10 µM), Fe deficiency, or Mn deficiency. Data are mean CsMTP6 transcript levels (±SD) relative to the constitutively expressed reference gene CACS calculated from the arithmetic means of ?Cp values obtained in three independent experiments. (B) Western blot analysis of CsMTP6 protein levels in mitochondria isolated from roots of cucumbers grown under different Fe and Mn treatments for 2 weeks. Immunoblots of the same mitochondria were probed with antibodies against mitochondrial COXII cytochrome oxidase to ensure equal loading of membrane protein (10 µg) onto SDS-PAGE. Significant differences were determined using Tukey’s test.


    Reactant: Arabidopsis thaliana (Thale cress)

    Application: Western Blotting

    Pudmed ID: 32457324

    Journal: Sci Rep

    Figure Number: 4C

    Published Date: 2020-05-26

    First Author: Sako, K., Futamura, Y., et al.

    Impact Factor: 4.13

    Open Publication

    FSL0260 interacts with mitochondrial complex I and inhibits its activity. (a) Inhibition of complex I by FSL0260. Oxygen consumption rate (OCR) of isolated mitochondria from potato tuber was monitored with deamino-NADH in the absence or presence of FSL0260. (b) Inhibition of deamino-NADH oxidation by FSL0260. Deamino-NADH oxidation was measured by spectrometry using sonicated mitochondria isolated from the potato tuber. The experiment was conducted using three biological replicates. Error bars represent the mean ± SE (n?=?3). (c) Pull-down assay of FLS0260. Sonicated potato mitochondria were incubated with control or FSL0260 beads. Immunoblot assay was performed using an anti-gamma CA antibody and an anti-COX II antibody.

  • Additional Information
  • Additional information:

    Cellular [compartment marker] of mitochondrial inner membrane

    Additional information (application):
    Antibody detects COXII protein most optimally in membrane fractions. The signal is weak in a in total protein extract.

    Blue Native gel electrophoresis (BN-PAGE) has been performed on samples solubilized with digitonin (4:1) and loaded at 100 µg/well. Gel thickness was 2 mm with 4.5-16 % gradient.
  • Background
  • Background:

    Cytochrome c oxidase (COX) catalyzes the reduction of oxygen to water in the respiratory chain in the inner mitochondrial membrane. Subunits 1-3 form the functional core of the enzyme complex. Subunit 2 (COXII) transfers the electrons from cytochrome c via its binuclear copper A center to the bimetallic center of the catalytic subunit 1. Alternative name: cytochrome c oxidase subunit 2

  • Product Citations
  • Selected references: Xue et al.(2023). The PtdIns3P phosphatase MtMP promotes symbiotic nitrogen fixation via mitophagy in Medicago truncatula. iScience. 2023 Sep 15;26(10):107752.doi: 10.1016/j.isci.2023.107752.
    Hofmann, Wienkoop & Luthje (2022) Hypoxia-Induced Aquaporins and Regulation of Redox Homeostasis by a Trans-Plasma Membrane Electron Transport System in Maize Roots. Antioxidants (Basel). 2022 Apr 25;11(5):836. doi: 10.3390/antiox11050836. PMID: 35624700; PMCID: PMC9137787.
    Kumar et al. (2022). Proteomic dissection of rice cytoskeleton reveals the dominance of microtubule and microfilament proteins, and novel components in the cytoskeleton-bound polysome, Plant Physiology and Biochemistry, Volume 170,2022,Pages 75-86,ISSN 0981-9428, https://doi.org/10.1016/j.plaphy.2021.11.037.
    Pavlovic & Kocab. (2021) Alternative oxidase (AOX) in the carnivorous pitcher plants of the genus Nepenthes: what is it good for? Ann Bot. 2021 Dec 18:mcab151. doi: 10.1093/aob/mcab151. Epub ahead of print. PMID: 34922341.
    Makino et al. (2020). Induction of Terminal Oxidases of Electron Transport Chain in Broccoli Heads Under Controlled Atmosphere Storage. Foods, 9 (4)
    Wang et al. (2020) Rerouting of ribosomal proteins into splicing in plant organelles. BioRxiv, DOI: 10.1101/2020.03.03.974766 .
  • Protocols
  • Preparation of plant mitochondrial fraction

    Agrisera Western Blot protocol and video tutorials

    Protocols to work with plant and algal protein extracts
  • Reviews:
  • Reviews
    Below, you can grade the product on a scale from 0 to 5.
    Please also provide information about species, application, dilution and obtained result for the reviewed antibody.
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    Number of reviews: (5)
    Jane Lee | 2023-12-07
    0 | 0
    This antibody works very well for our samples.
    G.Vanlerberghe | 2018-02-21
    0 | 0
    This antibody worked effectively in Nicotiana tabacum, using isolated leaf mitochondria.
    Michal RUREK | 2014-02-14
    0 | 0
    10 g of mitochondrial proteins from cauliflower (Brassica oleracea var. botrytis) curds (apical layer) was separated on 12 % SDS-PAGE (Laemmli- type) and blotted 1h to Immobilone P (Millipore) using Sedryt apparatus (Kucharczyk). After blocking (5% milk in PBS-T) for 1 h at RT, blots were incubated in the primary antibody at a dilution of 1: 1000 for overnight at 4 deg. with agitation. The primary antibody, diluted in 2% milk in PBS-T was reused several times. Secondary, anti-rabbit HRP- linked antibodies were bound at 1/10000 dilution at RT (1h). Single band of ca. 30 kDa was detected using standart GE Healthcare ECL reagents.
    Gerhard Obermeyer | 2009-11-18
    0 | 0
    The antibody worked well in Western blots and gave a main signal at ca. 30 kDa in a microsomal fraction prepared from Lilium pollen grains. Anti-COXII was diluted 1:1,000 and an anti-chicken IgY HRP-conjugated (1:10,000) was used for detection.
    David Macherel | 2009-07-23
    0 | 0
    The antibody yields a clean signal on western blots with extracts from germinating Arabidopsis seeds, young seedlings and leaves. We use it at 1/1000 dilution followed by ECL detection

Accessories

AS04 052  |  Clonality: Polyclonal  |  Host: Hen  |  Reactivity: A. thaliana (leaf extract and isolated mitochondria), B. oleracea, P. major, P. euryphylla, S. uniflora, S. dioica

416 €
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AS09 602 |  Clonality: Polyclonal | Host: Goat | Reactivity: Rabbit IgG (H&L)

213 €
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