MA7-00308-A488

antibody from Invitrogen Antibodies

Targeting: EGFR ERBB, ERBB1

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA7-00308-A488

Provider

Invitrogen Antibodies

Product name

EGFR Recombinant Rabbit Monoclonal Antibody (30H45L48), Alexa Fluor™ 488

Antibody type

Monoclonal

Antigen

Other

Description

This antibody is predicted to react with canine, mouse and rat based on sequence homology.

Conjugate

Green dye

Antibody clone number

30H45L48

Concentration

1 mg/mL

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of EGFR was performed using 90% confluent log phase A-431 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with EGFR Alexa Fluor 488 Recombinant Rabbit Monoclonal Antibody (Product # MA7-00308-A488) at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing membrane localization. Panel e represents the isotype control. The images were captured at 60X magnification.

Conjugate

Green dye

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of EGFR (green) in HeLa cells. The cells were fixed with 4% Paraformaldehyde in PBS for 15 minutes at room temperature, and blocked with 3% BSA in PBS (Product # 37525) for 30 minutes at room temperature. Cells were stained with a EGFR Monoclonal Antibody, AlexaFluor 488 conjugate (Product # MA7-00308-A488) at a dilution of 2.5 µg/mL in blocking buffer for 1 hour at room temperature protected from light. Nuclei (blue) were stained with Hoechst Dye (Product # 62249) at a dilution of 1:10,000 in blocking buffer. Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.

Conjugate

Green dye

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of EGFR was performed using 70% confluent log phase A-431 cells (WIld type, panels a,d), CAS9 control (panels b,e) and EGFR Knockout (panels c,f). The cells were fixed, permeabilized, and labelled with EGFR, Alexa Fluor 488 (30H45L48), ABfinity™ Rabbit Monoclonal Antibody(Product # MA7-00308-A488, 5 µg/mL). Nuclei (blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938) and Rhodamine Phalloidin (Product # R415, 1:300) was used for cytoskeletal F-actin (red) staining. Loss of signal was observed in EGFR Knockout cells (panel c,f) confirming specificity of the antibody to EGFR(green). The images were captured at 60X magnification.

Conjugate

Green dye

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of EGFR-Alexa Fluor® 488 conjugate was achieved by transfecting A-431 cells with EGFR specific siRNA (Silencer® select Cat # s563+s564+s565). Immunofluorescence analysis was performed on A-431 cells (untransfected, panel a,d), transfected with non-specific scrambled siRNA (panels b,e) and transfected with EGFR specific siRNA (panel c,f). Cells were fixed, permeabilized, and labelled with EGFR-Alexa Fluor® 488 conjugate Rabbit monoclonal Antibody (Product # MA7-00308-A488, 5 µg/mL), Nuclei (blue) were stained using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938), and Rhodamine Phalloidin (Product # R415, 1:300) was used for cytoskeletal F-actin (red) staining. Reduction of signal was observed upon siRNA mediated knockdown (panel c,f) confirming specificity of the antibody to EGFR(green). The images were captured at 60X magnification.

Conjugate

Green dye