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MCU Antibody (YA704)

(Synonyms: C10orf42, CCDC109A, MCU, HsMCU, Coiled-coil domain-containing protein 109A)
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Based on 1 publication(s) in Google Scholar

MCU Antibody (YA704) is a Mouse-derived and non-conjugated IgM monoclonal antibody, targeting to MCU.

For research use only. We do not sell to patients.
  • Host:

    Mouse

  • Isotype:

    IgM

  • Application:

    WB, IHC-P, ICC/IF

  • Reactivity :

    Human, Mouse, Rat, Monkey

  • Formulation:

    1.Supplied in PBS (pH7.4), 0.2% BSA and 50% Glycerol. Preservative: 0.05% Sodium Azide.
    2.Supplied in PBS (pH7.4), 0.1% BSA and 40% Glycerol. Preservative: 0.05% Sodium Azide.
    Please refer to the lot-specific COA for specific buffer information.

  • Conjugation:
    Non-conjugated
  • Experimental Validation Results for MCU Antibody (YA704)
  • Experimental Validation Results for MCU Antibody (YA704)
  • MCU Antibody (YA704) Product Image
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Applications

Application
WB Info
WB: Western Blot
IHC-P Info
IHC-P: Immunohistochemistry-Paraffin
IF/ICC Info
 
Dilution Ratio 1:5000 1:50 1:50

Product Details

Description

MCU Antibody (YA704) is a Mouse-derived and non-conjugated IgM monoclonal antibody, targeting to MCU.

  • Host Mouse
  • Clonality Monoclonal
  • Species Reactivity
    Human, Mouse, Rat, Monkey
  • Observed Molecular Weight
    Observed band size: 35 kDa Info
    Note: Due to possible protein modifications or aggregation, the molecular weight should be confirmed by actual measurement, and the predicted value is for reference only.
  • Calculated Molecular Weight Predicted band size: 40 kDa,
Species Reactivity Database

Entrez Gene: 90550 Human

SwissProt: Q8NE86 Human

Immunogen

Synthetic peptide corresponding to Human MCU.AA range:51-325 / 351.

Sensitivity

Endogenous

Purification

Protein G affinity purified.

Conjugation

Non-conjugated

Modification

Unmodified

Isotype

IgM

RRID

AB_3102354

Product Properties

  • Appearance

    Solution

  • Formulation

    1.Supplied in PBS (pH7.4), 0.2% BSA and 50% Glycerol. Preservative: 0.05% Sodium Azide.
    2.Supplied in PBS (pH7.4), 0.1% BSA and 40% Glycerol. Preservative: 0.05% Sodium Azide.
    Please refer to the lot-specific COA for specific buffer information.

  • Concentration

    Batch-dependent, Please check the COA for the concentration of each lot. Check Lot Concentration

  • Storage & Stability

    Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

  • Shipping

    Shipping with blue ice.

Verification Images

  • Experimental Validation Results for MCU Antibody (YA704)
    Western blot analysis was performed on protein extracts (25 μg) from 293T (lane 2), HepG2 (lane 3), HCT116 (lane 4), HeLa (lane 5) and Jurkat (lane 6) using MCU antibody. Proteins were transferred onto a 0.45 μm PVDF membrane using the Trans-Blot® Turbo™ system for 13 min. The membrane was then blocked with 5% nonfat milk in TBST (HY-K1025) for 1 h at room temperature. The primary antibody (1:1000) and loading control antibody GAPDH Antibody (HRP) (HY-P80954A) (1:5000) were diluted in 5% nonfat milk in TBST and incubated with the membrane overnight at 4°C. After washing, the membrane of primary antibody was incubated with HRP-conjugated goat anti-rabbit/mouse IgG secondary antibody (HY-P8001/HY-P8004) (1:5000) diluted in 5% nonfat milk in TBST for 1 h at room temperature. Protein bands were visualized using an Ultra High Sensitivity ECL detection kit (HY-K1005).
  • Experimental Validation Results for MCU Antibody (YA704)
    Western blot analysis of extracts from HEK293(lane 2(20μg) or lane 3(40μg)) and HepG2(lane 4(20μg) or lane 5(40μg)), using MCU (HY-P80216) Mouse mAb. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody (HY-P80216, 1/1000) and Loading control antibody (GAPDH, HY-P80954, 1/3000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (HY-P8004/HY-P8001, 1/10,000) was used for 1 hour at room temperature.

Background

  • Function

    MCU is a Channel-forming and calcium-conducting subunit of the mitochondrial inner membrane calcium uniporter complex (uniplex), which mediates calcium uptake into the mitochondrial matrix. MCU channel activity is regulated by the calcium-sensor subunits of the uniplex MICU1 and MICU2 (or MICU3). Mitochondrial calcium homeostasis plays key roles in cellular physiology and regulates ATP production, cytoplasmic calcium signals and activation of cell death pathways. Involved in buffering the amplitude of systolic calcium rises in cardiomyocytes. While dispensable for baseline homeostatic cardiac function, acts as a key regulator of short-term mitochondrial calcium loading underlying a 'fight-or-flight' response during acute stress: acts by mediating a rapid increase of mitochondrial calcium in pacemaker cells. Participates in mitochondrial permeability transition during ischemia-reperfusion injury. Mitochondrial calcium uptake in skeletal muscle cells is involved in muscle size in adults. Regulates synaptic vesicle endocytosis kinetics in central nerve terminal. Regulates glucose-dependent insulin secretion in pancreatic beta-cells by regulating mitochondrial calcium uptake. Involved in antigen processing and presentation[1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20][21][22][23].

  • Subcellular Localization

    Mitochondrion inner membrane; Multi-pass membrane protein

  • Expression


    Induction:MCU transcripts are down-regulated by microRNA miR-25 (PubMed:23246404) . Down-regulation by miR-25 may protect cardiomyocytes against oxidative damage in cardiomyocytes (PubMed:25764156)

  • Isoforms & Post-Translational Modification

    Q8NE86 has 3 isomers: Q8NE86-1: 39867 Da (predicted); Q8NE86-2: 36996 Da (predicted); Q8NE86-3: 35159 Da (predicted).
    Phosphorylation by CaMK2 in heart leads to increased MCU current (PubMed:23051746, PubMed:25254481). The regulation of MCU by CaMK2 is however subject to discussion: another group was unable to reproduce these results (PubMed:25254480). Phosphorylated on tyrosines by PTK2B/PYK2, promoting oligomerization (PubMed:24800979);Glutathionylation at Cys-97 in response to reactive oxygen species (ROS) promotes MCU higher-order assembly, leading to constitutive activation of the MCU channel and mitochondrial calcium overload;Undergoes proteolytic degradation by SPG7

  • Subunit

    Homotetramer (PubMed:32494073). Component of the uniplex complex, composed of MCU, EMRE/SMDT1, MICU1 and MICU2 (or MICU3) in a 4:4:1:1 stoichiometry (PubMed:24231807, PubMed:26341627, PubMed:31080062, PubMed:32494073, PubMed:32862359). Interacts with CCDC109B/MCUB; this inhibits channel activity (PubMed:27184846). Interacts with MCUR1 (PubMed:23178883, PubMed:26341627, PubMed:26976564, PubMed:27184846). Interactions with MICU1 and MCUR1 are mutually exclusive (PubMed:23178883). Interacts with SLC25A23 (PubMed:24430870)

  • SwissProt ID

    Q8NE86

  • Gene ID
    90550 [NCBI]
  • Synonyms

    C10orf42, CCDC109A, MCU, HsMCU, Coiled-coil domain-containing protein 109A

  • Research Field

    Cardiovascular

[1]. Baughman JM, et al. Integrative genomics identifies MCU as an essential component of the mitochondrial calcium uniporter. Nature. 2011 Jun 19;476(7360):341-5. [Content Brief]

[2]. De Stefani D, et al. A forty-kilodalton protein of the inner membrane is the mitochondrial calcium uniporter. Nature. 2011 Jun 19;476(7360):336-40. [Content Brief]

[3]. Drago I, et al. Mitochondrial Ca2+ uptake contributes to buffering cytoplasmic Ca2+ peaks in cardiomyocytes. Proc Natl Acad Sci U S A. 2012 Aug 7;109(32):12986-91. [Content Brief]

[4]. Tarasov AI, et al. The mitochondrial Ca2+ uniporter MCU is essential for glucose-induced ATP increases in pancreatic β-cells. PLoS One. 2012;7(7):e39722. [Content Brief]

[5]. Alam MR, et al. Mitochondrial Ca2+ uptake 1 (MICU1) and mitochondrial ca2+ uniporter (MCU) contribute to metabolism-secretion coupling in clonal pancreatic β-cells. J Biol Chem. 2012 Oct 5;287(41):34445-54. [Content Brief]

[6]. Mallilankaraman K, et al. MICU1 is an essential gatekeeper for MCU-mediated mitochondrial Ca(2+) uptake that regulates cell survival. Cell. 2012 Oct 26;151(3):630-44. [Content Brief]

[7]. Mallilankaraman K, et al. MCUR1 is an essential component of mitochondrial Ca2+ uptake that regulates cellular metabolism. Nat Cell Biol. 2012 Dec;14(12):1336-43. [Content Brief]

[8]. Chaudhuri D, et al. MCU encodes the pore conducting mitochondrial calcium currents. Elife. 2013 Jun 4;2:e00704. [Content Brief]

[9]. Hoffman NE, et al. MICU1 motifs define mitochondrial calcium uniporter binding and activity. Cell Rep. 2013 Dec 26;5(6):1576-1588. [Content Brief]

[10]. Patron M, et al. MICU1 and MICU2 finely tune the mitochondrial Ca2+ uniporter by exerting opposite effects on MCU activity. Mol Cell. 2014 Mar 6;53(5):726-37. [Content Brief]

[11]. Lee Y, et al. Structure and function of the N-terminal domain of the human mitochondrial calcium uniporter. EMBO Rep. 2015 Oct;16(10):1318-33. [Content Brief]

[12]. Yoo J, et al. Cryo-EM structure of a mitochondrial calcium uniporter. Science. 2018 Aug 3;361(6401):506-511. [Content Brief]

[13]. Baradaran R, et al. Cryo-EM structures of fungal and metazoan mitochondrial calcium uniporters. Nature. 2018 Jul;559(7715):580-584. [Content Brief]

[14]. Paillard M, et al. MICU1 Interacts with the D-Ring of the MCU Pore to Control Its Ca(2+) Flux and Sensitivity to Ru360. Mol Cell. 2018 Nov 15;72(4):778-785.e3. [Content Brief]

[15]. Phillips CB, et al. The conserved aspartate ring of MCU mediates MICU1 binding and regulation in the mitochondrial calcium uniporter complex. Elife. 2019 Jan 15;8:. [Content Brief]

[16]. Wang Y, et al. Structural Mechanism of EMRE-Dependent Gating of the Human Mitochondrial Calcium Uniporter. Cell. 2019 May 16;177(5):1252-1261.e13. [Content Brief]

[17]. Fan M, et al. Structure and mechanism of the mitochondrial Ca(2+) uniporter holocomplex. Nature. 2020 Jun;582(7810):129-133. [Content Brief]

[18]. Wang Y, et al. Structural insights into the Ca(2+)-dependent gating of the human mitochondrial calcium uniporter. Elife. 2020 Aug 7;9:. [Content Brief]

[19]. Van Keuren AM, et al. Mechanisms of EMRE-Dependent MCU Opening in the Mitochondrial Calcium Uniporter Complex. Cell Rep. 2020 Dec 8;33(10):108486. [Content Brief]

[20]. Tsai CW, et al. Evidence supporting the MICU1 occlusion mechanism and against the potentiation model in the mitochondrial calcium uniporter complex. Proc Natl Acad Sci U S A. 2023 Apr 18;120(16):e2217665120. [Content Brief]

[21]. Rodríguez-Prados M, et al. MICU1 occludes the mitochondrial calcium uniporter in divalent-free conditions. Proc Natl Acad Sci U S A. 2023 May 9;120(19):e2218999120. [Content Brief]

[22]. Matesanz-Isabel J, et al. Functional roles of MICU1 and MICU2 in mitochondrial Ca(2+) uptake. Biochim Biophys Acta. 2016 Jun;1858(6):1110-7. [Content Brief]

[23]. Wu Y, et al. The mitochondrial uniporter controls fight or flight heart rate increases. Nat Commun. 2015 Jan 20;6:6081. [Content Brief]

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Measured by its ability to inhibit anti-CD3-induced proliferation of stimulated CTLL-2 mouse cytotoxic T cells. The ED50 this effect is 0.8023 μg/ml, corresponding to a specific activity is 1246.42 units/mg.

Assay Types
  • 1/16

    MedChemExpress Validation
     Western blot analysis was performed on protein extracts (25 μg) from 293T (lane 2), HepG2 (lane 3), HCT116 (lane 4), HeLa (lane 5) and Jurkat (lane 6) using MCU antibody. Proteins were transferred onto a 0.45 μm PVDF membrane using the Trans-Blot® Turbo™ system for 13 min. The membrane was then blocked with 5% nonfat milk in TBST (HY-K1025) for 1 h at room temperature. The primary antibody (1:1000) and loading control antibody GAPDH Antibody (HRP) (HY-P80954A) (1:5000) were diluted in 5% nonfat milk in TBST and incubated with the membrane overnight at 4°C. After washing, the membrane of primary antibody was incubated with HRP-conjugated goat anti-rabbit/mouse IgG secondary antibody (HY-P8001/HY-P8004) (1:5000) diluted in 5% nonfat milk in TBST for 1 h at room temperature. Protein bands were visualized using an Ultra High Sensitivity ECL detection kit (HY-K1005).

  • 1/16

    MedchemExpress Validation
     Western blot analysis of extracts from HEK293(lane 2(20μg) or lane 3(40μg)) and HepG2(lane 4(20μg) or lane 5(40μg)), using MCU (HY-P80216) Mouse mAb. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody (HY-P80216, 1/1000) and Loading control antibody (GAPDH, HY-P80954, 1/3000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (HY-P8004/HY-P8001, 1/10,000) was used for 1 hour at room temperature.

  • MCU Antibody (YA704) Product Image
    1/16
  • Experimental Validation Results for MCU Antibody (YA704)
  • Experimental Validation Results for MCU Antibody (YA704)
  • MCU Antibody (YA704) Product Image
  • Experimental Validation Results for MCU Antibody (YA704)
    1/1
    MedChemExpress Validation
     Western blot analysis was performed on protein extracts (25 μg) from 293T (lane 2), HepG2 (lane 3), HCT116 (lane 4), HeLa (lane 5) and Jurkat (lane 6) using MCU antibody. Proteins were transferred onto a 0.45 μm PVDF membrane using the Trans-Blot® Turbo™ system for 13 min. The membrane was then blocked with 5% nonfat milk in TBST (HY-K1025) for 1 h at room temperature. The primary antibody (1:1000) and loading control antibody GAPDH Antibody (HRP) (HY-P80954A) (1:5000) were diluted in 5% nonfat milk in TBST and incubated with the membrane overnight at 4°C. After washing, the membrane of primary antibody was incubated with HRP-conjugated goat anti-rabbit/mouse IgG secondary antibody (HY-P8001/HY-P8004) (1:5000) diluted in 5% nonfat milk in TBST for 1 h at room temperature. Protein bands were visualized using an Ultra High Sensitivity ECL detection kit (HY-K1005).
  • Experimental Validation Results for MCU Antibody (YA704)
    1/1
    MedchemExpress Validation
     Western blot analysis of extracts from HEK293(lane 2(20μg) or lane 3(40μg)) and HepG2(lane 4(20μg) or lane 5(40μg)), using MCU (HY-P80216) Mouse mAb. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody (HY-P80216, 1/1000) and Loading control antibody (GAPDH, HY-P80954, 1/3000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (HY-P8004/HY-P8001, 1/10,000) was used for 1 hour at room temperature.
  • Experimental Validation Results for MCU Antibody (YA704)
  • Experimental Validation Results for MCU Antibody (YA704)
  • MCU Antibody (YA704) Product Image
    MedChemExpress Validation

    Western blot analysis was performed on protein extracts (25 μg) from 293T (lane 2), HepG2 (lane 3), HCT116 (lane 4), HeLa (lane 5) and Jurkat (lane 6) using MCU antibody. Proteins were transferred onto a 0.45 μm PVDF membrane using the Trans-Blot® Turbo™ system for 13 min. The membrane was then blocked with 5% nonfat milk in TBST (HY-K1025) for 1 h at room temperature. The primary antibody (1:1000) and loading control antibody GAPDH Antibody (HRP) (HY-P80954A) (1:5000) were diluted in 5% nonfat milk in TBST and incubated with the membrane overnight at 4°C. After washing, the membrane of primary antibody was incubated with HRP-conjugated goat anti-rabbit/mouse IgG secondary antibody (HY-P8001/HY-P8004) (1:5000) diluted in 5% nonfat milk in TBST for 1 h at room temperature. Protein bands were visualized using an Ultra High Sensitivity ECL detection kit (HY-K1005).

  • MCU Antibody (YA704) Product Image
    MedchemExpress Validation

    Western blot analysis of extracts from HEK293(lane 2(20μg) or lane 3(40μg)) and HepG2(lane 4(20μg) or lane 5(40μg)), using MCU (HY-P80216) Mouse mAb. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody (HY-P80216, 1/1000) and Loading control antibody (GAPDH, HY-P80954, 1/3000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (HY-P8004/HY-P8001, 1/10,000) was used for 1 hour at room temperature.

  • MCU Antibody (YA704) Product Image WB
  • MCU Antibody (YA704) Product Image WB