Anti-MG53 antibody [EPR26057-7]

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-MG53 antibody [EPR26057-7] (AB307593)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat cerebrum (fresh) tissue labeling MG53 with ab307593 at 1/100 dilution (5.15 µg/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 µg/mL) (Green). Negative control : confocal image showing no staining on rat cerebrum (PMID : 20139895). The nuclear counterstain was DAPI (Blue). The section was incubated with ab307593 for 60 mins at room temperature. The section was then mounted using Fluoromount®. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 µg/mL).

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-MG53 antibody [EPR26057-7] (AB307593)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse cerebrum (fresh) tissue labeling MG53 with ab307593 at 1/100 dilution (5.15 µg/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 µg/mL) (Green). Negative control : confocal image showing no staining on mouse cerebrum (PMID : 20139895). The nuclear counterstain was DAPI (Blue). The section was incubated with ab307593 for 60 mins at room temperature. The section was then mounted using Fluoromount®. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 µg/mL).

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-MG53 antibody [EPR26057-7] (AB307593)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat skeletal muscle (fresh) tissue labeling MG53 with ab307593 at 1/100 dilution (5.15 µg/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 µg/mL) (Green). Confocal image showing positive staining on rat skeletal muscle. The nuclear counterstain was DAPI (Blue). The section was incubated with ab307593 for 60 mins at room temperature. The section was then mounted using Fluoromount®. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 µg/mL).

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-MG53 antibody [EPR26057-7] (AB307593)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse skeletal muscle (fresh) tissue labeling MG53 with ab307593 at 1/100 dilution (5.15 µg/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 µg/mL) (Green). Confocal image showing positive staining on mouse skeletal muscle. The nuclear counterstain was DAPI (Blue). The section was incubated with ab307593 for 60 mins at room temperature. The section was then mounted using Fluoromount®. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 µg/mL).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-MG53 antibody [EPR26057-7] (AB307593)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 100% methanol-permeabilized C2C12 (mouse myoblast) cells labeling MG53 with ab307593 at 1/500 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing membranous and cytoplasmic staining in differentiated C2C12 cells and showing no staining in undifferentiated C2C12 cells. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (2.5 µg/ml) (Red). Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• IP

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Immunoprecipitation - Anti-MG53 antibody [EPR26057-7] (AB307593)

MG53 was immunoprecipitated from 0.35 mg C2C12 (differentiation for 7 days) whole cell lysate 5 µg with ab307593 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307593 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : C2C12 (differentiation for 7 days) whole cell lysate 5 µg Lane 2 : ab307593 IP in (C2C12 differentiation for 7 days) whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab307593 in C2C12 (differentiation for 7 days) whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 8 seconds.

All lanes:

Immunoprecipitation - Anti-MG53 antibody [EPR26057-7] (ab307593) at 1/30 dilution

All lanes:

C2C12

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 8s

• WB

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Western blot - Anti-MG53 antibody [EPR26057-7] (AB307593)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. Negative control : kidney (PMID : 20139895), brain (PMID : 20139895). Exposure time : 37 seconds.

All lanes:

Western blot - Anti-MG53 antibody [EPR26057-7] (ab307593) at 1/1000 dilution

Lane 1:

Human skeletal muscle tissue lysate at 20 µg

Lane 2:

Human kidney tissue lysate at 20 µg

Lane 3:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 4:

Mouse heart tissue lysate at 20 µg

Lane 5:

Mouse brain tissue lysate at 20 µg

Lane 6:

Mouse kidney tissue lysate at 20 µg

Lane 7:

Rat skeletal muscle tissue lysate at 20 µg

Lane 8:

Rat brain tissue lysate at 20 µg

Lane 9:

Rat kidney tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 53 kDa

false

• WB

Supplier Data

Western blot - Anti-MG53 antibody [EPR26057-7] (AB307593)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. Negative control : MCF7 (Human Protein Atlas database). Exposure time : 37 seconds.

All lanes:

Western blot - Anti-MG53 antibody [EPR26057-7] (ab307593) at 1/1000 dilution

Lane 1:

HCT116 (human colorectal carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

Untreated C2C12 (mouse myoblasts myoblast) whole cell lysate at 20 µg

Lane 4:

C2C12 differentiation for 7 days whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 53 kDa

false

Exposure time: 37s