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MCU Mouse Monoclonal Antibody [A2E11]

Usd: 350

Specification

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Catalog# EM1901-85

MCU Mouse Monoclonal Antibody [A2E11]

Application

  • WB

  • IHC-P

  • IF-Cell

Reactivity

  • Human

  • Mouse

  • Rat

  • Green monkey

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

MCU Mouse Monoclonal Antibody [A2E11]

Antibody Type

Mouse Monoclonal Antibody

Immunogen

Recombinant protein within Human MCU aa 51-325 / 351.

Species Reactivity

Human, Mouse, Rat, Green monkey

Validated Applications

WB, IHC-P, IF-Cell

Molecular Weight

Predicted band size: 40 kDa

Positive Control

HCT 116 cell lysate, PANC-1 cell lysate, A549 cell lysate, HeLa cell lysate, COS-1 cell lysate, NIH/3T3 cell lysate, C2C12 cell lysate, C6 cell lysate, PC-12 cell lysate, Mouse colon tissue lysate, Mouse testis tissue lysate, Rat colon tissue lysate, Rat heart tissue lysate, human colon tissue, human colon cancer tissue, HCT 116, NIH/3T3, C6.

Conjugation

unconjugated

Clone Number

A2E11

RRID

AB_3068930

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgM

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:5,000

  • IHC-P

  • 1:50

  • IF-Cell

  • 1:50

Target

Function

The mitochondrial calcium uniporter (MCU) is a transmembrane protein that allows the passage of calcium ions from a cell's cytosol into mitochondria. Its activity is regulated by MICU1 and MICU2, which together with the MCU make up the mitochondrial calcium uniporter complex. The MCU is one of the primary sources of mitochondria uptake of calcium, and flow is dependent on membrane potential of the inner mitochondrial membrane and the concentration of calcium in the cytosol relative to the concentration in the mitochondria. Balancing calcium concentration is necessary to increase the cell's energy supply and regulate cell death. Calcium is balanced through the MCU in conjunction with the sodium-calcium exchanger. The MCU has a very low affinity for calcium, so the cytosolic calcium concentration needs to be approximately 5-10 uM for significant transport of calcium into the mitochondria. Mitochondria are closely associated with the endoplasmic reticulum (ER), at contact sites, which contains stores of cellular calcium ions for calcium signaling. The presence of 1,4,5-triphosphate (IP3) triggers the release of calcium from these intracellular stores, which creates microdomains of high calcium concentration between the ER and the mitochondria, creating the conditions for the MCU to take up calcium.

Background References

1. Wang P et al. Elevated MCU Expression by CaMKIIdeltaB Limits Pathological Cardiac Remodeling. Circulation. 2022 Apr

2. Li C et al. Implications of MCU complex in metabolic diseases. FASEB J. 2023 Aug

Sequence Similarity

Belongs to the MCU (TC 1.A.77) family.

Post-translational Modification

Phosphorylation by CaMK2 in heart leads to increased MCU current. The regulation of MCU by CaMK2 is however subject to discussion: another group was unable to reproduce these results. Phosphorylated on tyrosines by PTK2B/PYK2, promoting oligomerization.

Subcellular Location

Mitochondrion inner membrane.

UNIPROT

SwissProt: Q8NE86 Human

SwissProt: Q3UMR5 Mouse

Entrez Gene: 294560 Rat

Synonyms

C109A_HUMAN antibody

C10orf42 antibody

Calcium uniporter protein mitochondrial antibody

Ccdc109a antibody

Coiled-coil domain-containing protein 109A antibody

HsMCU antibody

Mitochondrial Calcium Uniporter antibody

Images

  • Western blot analysis of MCU on different lysates with Mouse anti-MCU antibody (<a href="/products/EM1901-85" style="font-weight: bold;text-decoration: underline;">EM1901-85</a>) at 1/5,000 dilution.<br /><br />Lane 1: HCT 116 cell lysate (20 µg/Lane)<br />Lane 2: PANC-1 cell lysate (20 µg/Lane)<br />Lane 3: A549 cell lysate (20 µg/Lane)<br />Lane 4: HeLa cell lysate (20 µg/Lane)<br />Lane 5: COS-1 cell lysate (20 µg/Lane)<br />Lane 6: NIH/3T3 cell lysate (20 µg/Lane)<br />Lane 7: C2C12 cell lysate (20 µg/Lane)<br />Lane 8: C6 cell lysate (20 µg/Lane)<br />Lane 9: PC-12 cell lysate (20 µg/Lane)<br />Lane 10: Mouse colon tissue lysate (20 µg/Lane)<br />Lane 11: Mouse testis tissue lysate (20 µg/Lane)<br />Lane 12: Rat colon tissue lysate (20 µg/Lane)<br />Lane 13: Rat heart tissue lysate (20 µg/Lane)<br /><br />Predicted band size: 40 kDa<br />Observed band size: 33 kDa<br /><br />Exposure time: 59 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/EM1901-85" style="font-weight: bold;text-decoration: underline;">EM1901-85</a>) at 1/5,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of MCU on different lysates with Mouse anti-MCU antibody (EM1901-85) at 1/5,000 dilution.

    Lane 1: HCT 116 cell lysate (20 µg/Lane)
    Lane 2: PANC-1 cell lysate (20 µg/Lane)
    Lane 3: A549 cell lysate (20 µg/Lane)
    Lane 4: HeLa cell lysate (20 µg/Lane)
    Lane 5: COS-1 cell lysate (20 µg/Lane)
    Lane 6: NIH/3T3 cell lysate (20 µg/Lane)
    Lane 7: C2C12 cell lysate (20 µg/Lane)
    Lane 8: C6 cell lysate (20 µg/Lane)
    Lane 9: PC-12 cell lysate (20 µg/Lane)
    Lane 10: Mouse colon tissue lysate (20 µg/Lane)
    Lane 11: Mouse testis tissue lysate (20 µg/Lane)
    Lane 12: Rat colon tissue lysate (20 µg/Lane)
    Lane 13: Rat heart tissue lysate (20 µg/Lane)

    Predicted band size: 40 kDa
    Observed band size: 33 kDa

    Exposure time: 59 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM1901-85) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human colon tissue with Mouse anti-MCU antibody (<a href="/products/EM1901-85" style="font-weight: bold;text-decoration: underline;">EM1901-85</a>) at 1/50 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/EM1901-85" style="font-weight: bold;text-decoration: underline;">EM1901-85</a>) at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human colon tissue with Mouse anti-MCU antibody (EM1901-85) at 1/50 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-85) at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Mouse anti-MCU antibody (<a href="/products/EM1901-85" style="font-weight: bold;text-decoration: underline;">EM1901-85</a>) at 1/50 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/EM1901-85" style="font-weight: bold;text-decoration: underline;">EM1901-85</a>) at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Mouse anti-MCU antibody (EM1901-85) at 1/50 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-85) at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunocytochemistry analysis of HCT 116 cells labeling MCU with Mouse anti-MCU antibody (<a href="/products/EM1901-85" style="font-weight: bold;text-decoration: underline;">EM1901-85</a>) at 1/50 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-MCU antibody (<a href="/products/EM1901-85" style="font-weight: bold;text-decoration: underline;">EM1901-85</a>) at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. beta Tubulin (<a href="/products/ET1602-4" style="font-weight: bold;text-decoration: underline;">ET1602-4</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 594, <a href="/products/HA1122" style="font-weight: bold;text-decoration: underline;">HA1122</a>) were used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HCT 116 cells labeling MCU with Mouse anti-MCU antibody (EM1901-85) at 1/50 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-MCU antibody (EM1901-85) at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

  • Immunocytochemistry analysis of NIH/3T3 cells labeling MCU with Mouse anti-MCU antibody (<a href="/products/EM1901-85" style="font-weight: bold;text-decoration: underline;">EM1901-85</a>) at 1/50 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-MCU antibody (<a href="/products/EM1901-85" style="font-weight: bold;text-decoration: underline;">EM1901-85</a>) at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. beta Tubulin (<a href="/products/ET1602-4" style="font-weight: bold;text-decoration: underline;">ET1602-4</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 594, <a href="/products/HA1122" style="font-weight: bold;text-decoration: underline;">HA1122</a>) were used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of NIH/3T3 cells labeling MCU with Mouse anti-MCU antibody (EM1901-85) at 1/50 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-MCU antibody (EM1901-85) at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

  • Immunocytochemistry analysis of C6 cells labeling MCU with Mouse anti-MCU antibody (<a href="/products/EM1901-85" style="font-weight: bold;text-decoration: underline;">EM1901-85</a>) at 1/50 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-MCU antibody (<a href="/products/EM1901-85" style="font-weight: bold;text-decoration: underline;">EM1901-85</a>) at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />beta Tubulin (<a href="/products/ET1602-4" style="font-weight: bold;text-decoration: underline;">ET1602-4</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 594, <a href="/products/HA1122" style="font-weight: bold;text-decoration: underline;">HA1122</a>) were used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of C6 cells labeling MCU with Mouse anti-MCU antibody (EM1901-85) at 1/50 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-MCU antibody (EM1901-85) at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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