MA5-13859

antibody from Invitrogen Antibodies

Targeting: EZR VIL2

Provider product page for MA5-13859

Western blot

Immunocytochemistry

Immunoprecipitation

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [42]
Comments [0]
Validations
Immunocytochemistry [5]
Immunohistochemistry [3]
Other assay [2]

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Product number: MA5-13859 - Provider product page
Provider: Invitrogen Antibodies
Product name: Anti-Ezrin Monoclonal Antibody (3C12), Biotin
Antibody type: Monoclonal
Antigen: Recombinant protein fragment
Description: MA5-13862 targets Ezrin in IHC (P), ICC, IP, IF, and WB applications and shows reactivity with Bovine, Hamster, Human, Marsupial, mouse, Non-human primate, and Rat samples. The MA5-13862 immunogen is a recombinant fragment of human ezrin corresponding to aa 362-585.
Reactivity: Human, Mouse, Rat, Bovine, Hamster
Host: Mouse
Conjugate: Biotin
Isotype: IgG
Antibody clone number: 3C12
Vial size: 500 µL
Concentration: 0.2 mg/ml
Storage: 4° C

EZR protein structure - MA5-13859 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry analysis of Ezrin in U2OS cells. Cells fixed with 4% paraformaldehyde were permeabilized with 0.1% Triton X-100 (Product # 28314) in PBS for 15 minutes at room temperature and blocked with 2% BSA in PBST (Product # 37525) for 30 minutes at room temperature. Cells were treated with Peroxidase Suppressor (Product # 35000), probed with an Ezrin monoclonal antibody (Product # MA5-13862) at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBST, and incubated with an HRP-conjugated goat anti-mouse IgG (H+L) secondary antibody (Product # 31431) at a dilution of 1:1000 for 30 minutes at room temperature. Chromogenic detection was performed using Metal Enhanced DAB Substrate Kit (Product # 34065). Images were taken on a Zeiss Axio Observer microscope at 20X magnification (x1.6 Optovar ~ 32X).
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry analysis of Ezrin in U2OS cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 (Product # 28314) in TBST (Product # 28376) for 30 minutes at room temperature and blocked with Universal Blocker (from Product # 03-6000) for 30 minutes at room temperature. Cells were probed with an Ezrin monoclonal antibody (Product # MA5-13862) at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBST, and incubated with an alkaline phosphatase-conjugated goat anti-mouse IgG (H+L) secondary antibody (Product # 31320) at a dilution of 1:250 for 30 minutes at room temperature. Chromogenic detection was performed using 1-Step NBT/BCIP (Product # 34042). Images were taken on a Zeiss Axio Observer microscope at 20X magnification (x1.6 Optovar ~ 32X).
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Phalloidin (orange) and Ezrin (green) in A431 cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in PBS for 10 minutes at room temperature and blocked with 2% BSA (Product # 37525) in PBS + 0.1% Triton X-100 for 30 minutes at room temperature. Cells were probed with an Ezrin monoclonal antibody (Product # MA5-13862) at a dilution of 1:75 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-mouse IgG secondary antibody (Product # 35502) at a dilution of 1:250 for 30 minutes at room temperature. Actin was stained with DyLight 550 Phalloidin (Product # 21835) at a dilution of 1:120 (2.5 units/ml final concentration) and nuclei (blue) were stained with Hoechst (Product # 62249) at a concentration of 1 µg/mL for 30 minutes. Images were taken on a Zeiss Axio Observer Z1 microscope at 20X magnification.
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Phalloidin (magenta) and Ezrin (green) in A431 cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in PBS for 10 minutes at room temperature and blocked with 2% BSA (Product # 37525) in PBS + 0.1% Triton X-100 for 30 minutes at room temperature. Cells were probed with an Ezrin monoclonal antibody (Product # MA5-13862) at a dilution of 1:75 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-mouse IgG secondary antibody (Product # 35502) at a dilution of 1:250 for 30 minutes at room temperature. Actin was stained with DyLight 633 Phalloidin (Product # 21840) at a dilution of 1:120 (2.5units/ml final concentration) and nuclei (blue) were stained with Hoechst (Product # 62249) at a concentration of 1 µg/mL for 30 minutes. Images were taken on a Zeiss Axio Observer Z1 microscope at 20X magnification.
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Phalloidin (dark red) and Ezrin (green) in A431 cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in PBS for 10 minutes at room temperature and blocked with 2% BSA (Product # 37525) in PBS + 0.1% Triton X-100 for 30 minutes at room temperature. Cells were probed with an Ezrin monoclonal antibody (Product # MA5-13862) at a dilution of 1:75 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-mouse IgG secondary antibody (Product # 35502) at a dilution of 1:250 for 30 minutes at room temperature. Actin was stained with DyLight 680 Phalloidin (Product # 21839) at a dilution of 1:120 (2.5 units/ml final concentration) and nuclei (blue) were stained with Hoechst (Product # 62249) at a concentration of 1 µg/mL for 30 minutes. Images were taken on a Zeiss Axio Observer Z1 microscope at 20X magnification.
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on paraffin-embedded human breast cancer tissue sections. To expose target proteins, heat-induced epitope retrieval was performed using 10mM sodium citrate (pH 6.0) buffer for 20 minutes at 95ºC. Following antigen retrieval, tissues were blocked in 3% BSA (Product # 37525) in PBST for 30 minutes at room temperature and then probed with an Ezrin monoclonal antibody (Product # MA5-13862) at a dilution of 1:100 for 1 hour in a humidified chamber. Tissues were washed extensively with PBS/0.025% Tween-20 (Product # 28314) and endogenous peroxidase activity quenched with Peroxidase Suppressor (Product # 35000) for 30 minutes at room temperature. Detection was performed using an HRP-conjugated goat anti-mouse IgG-HRP secondary antibody (Product # 31430) at a dilution of 1:500 followed by colorimetric detection using Metal Enhanced DAB Substrate Kit (Product # 34065). Images were taken on a Zeiss Axiovision microscope at 40X magnification.
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on paraffin-embedded human colon cancer tissue sections. To expose target proteins, heat-induced epitope retrieval was performed using 10mM sodium citrate (pH 6.0) buffer for 20 minutes at 95ºC. Following antigen retrieval, tissues were blocked in 3% BSA (Product # 37525) in PBST for 30 minutes at room temperature and then probed with an Ezrin monoclonal antibody (Product # MA5-13862) at a dilution of 1:100 for 1 hour in a humidified chamber. Tissues were washed extensively with PBS/0.025% Tween-20 (Product # 28314) and endogenous peroxidase activity quenched with Peroxidase Suppressor (Product # 35000) for 30 minutes at room temperature. Detection was performed using an HRP-conjugated goat anti-mouse IgG-HRP secondary antibody (Product # 31430) at a dilution of 1:500 followed by colorimetric detection using Metal Enhanced DAB Substrate Kit (Product # 34065). Images were taken on a Zeiss Axiovision microscope at 40X magnification.
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Formalin-fixed, paraffin-embedded human lung stained with Ezrin antibody using peroxidase-conjugate and DAB chromogen. Note cell membrane staining of bronchio-epithelial cells.
Conjugate: Biotin