MCU Mouse Monoclonal Antibody [A2E11]
cat.: EM1901-85
Product Type: Mouse monoclonal IgM, primary antibodies
Species reactivity: Human, Mouse, Rat, Monkey
Applications: WB, IHC-P, IF-Cell
Clonality: Monoclonal
Clone number: A2E11
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 40 kDa
Isotype: IgM
Immunogen: Recombinant protein within Human MCU aa 51-325 / 351.
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.
Subcellular location: Mitochondrion inner membrane.
Recommended Dilutions:
  WB
  IHC-P
  IF-Cell
1:5,000
1:50
1:50
Uniprot #: SwissProt: Q8NE86 Human | Q3UMR5 Mouse
Entrez Gene: 294560 Rat
Alternative names: C109A_HUMAN C10orf42 Calcium uniporter protein mitochondrial Ccdc109a Coiled-coil domain-containing protein 109A HsMCU Mitochondrial Calcium Uniporter
Images
EM1901-85_1.jpg Fig1: 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.
EM1901-85_2.jpg Fig2: 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.
EM1901-85_3.jpg Fig3: 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.
EM1901-85_4.jpg Fig4: 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.
EM1901-85_5.jpg Fig5: 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.
EM1901-85_6.jpg Fig6: 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.