For localisation studies, mESCs were plated in 0.1% gelatin (v/v) coated coverslips and fixed in 4% PFA in PBS for 20 min at room temperature (RT). Cells were permeabilised with a 0.5% Triton X-100 in PBS solution for 5 min at RT and then blocked with 1% Fish gelatin (w/v) in PBS solution for 30 min at RT. RLIM primary antibody (Novus Biologicals) was diluted 1:200 in blocking solution and added to cells for 2 h at RT. Anti-mouse Alexa-488 (Thermo Fisher Scientific) was used as a secondary antibody at 1:500 in blocking solution for 1 h at RT. Actin Red 555 reagent (Thermo Fisher Scientific, one drop per ml of blocking solution) was added together with secondary antibody for actin staining. Hoechst was added at 1:10,000 dilution in PBS for 5 min at RT as nuclear marker. Coverslips were mounted in glass slides using Fluorsave reagent (Millipore). Digital images were acquired in a Zeiss 710 confocal microscope and analysed and processed using ImageJ (NIH), Photoshop CC and Illustrator CC (Adobe).
Rlim primary antibody
Manufactured by Novus Biologicals
The RLIM primary antibody is a research-use only product designed to detect the RLIM protein in various biological samples. The RLIM protein is a regulatory protein involved in various cellular processes. This antibody can be utilized in techniques such as Western blotting and immunohistochemistry to study the expression and localization of the RLIM protein.
Automatically generated - may contain errors
2 protocols using rlim primary antibody
1
Immunofluorescence Localization of RLIM in mESCs
2
Immunocytochemistry of mESCs
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For localisation studies, mESCs were plated in 0.1% gelatin (v/v) coated coverslips and fixed in 4% PFA in PBS for 20 minutes at room temperature (RT). Cells were permeabilised with a 0.5% Triton X-100 in PBS solution for 5 minutes at RT and then blocked with 1% Fish gelatin (w/v) in PBS solution for 30 minutes at RT. RLIM primary antibody (Novus Biologicals) was diluted 1:200 in blocking solution and added to cells for 2 h at RT. Antimouse Alexa-488 (Thermo Fisher Scientific) was used as a secondary antibody at 1:500 in blocking solution for 1 h at RT. Actin Red 555 reagent (Thermo Fisher Scientific, one drop per ml of blocking solution) was added together with secondary antibody for actin staining.
Hoescht was added at 1:10000 dilution in PBS for 5 min at RT as nuclear marker. Coverslips were mounted in glass slides using Fluorsave reagent (Millipore). Digital images were acquired in a Zeiss 710 confocal microscope and analysed and processed using ImageJ (NIH), Photoshop CC and Illustrator CC (Adobe).
Hoescht was added at 1:10000 dilution in PBS for 5 min at RT as nuclear marker. Coverslips were mounted in glass slides using Fluorsave reagent (Millipore). Digital images were acquired in a Zeiss 710 confocal microscope and analysed and processed using ImageJ (NIH), Photoshop CC and Illustrator CC (Adobe).
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