711199

antibody from Invitrogen Antibodies

Targeting: RAF1 c-Raf, CRAF, Raf-1

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

711199

Provider

Invitrogen Antibodies

Product name

Phospho-c-Raf (Ser621) Recombinant Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

This antibody is predicted to react with Pig, Cat, Bovine, Sheep and Horse.

Concentration

0.5 mg/mL

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

For immunofluorescence analysis, SK-OV-3 cells were fixed and permeabilized for detection of endogenous Raf1 (pS621) using Raf1 (pS621) Recombinant Rabbit Polyclonal Antibody (Product # 711199, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of Raf1 (pS621) protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating nuclear localization of Raf1 (pS621). Panel e) shows loss of signal by competition with the Raf1 (pS621) phosphopeptide, demonstrating antibody specificity, and panel f) demonstrates no competition with the non-phospho peptide. Panel g) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.