Recombinant Anti-Fas antibody [EPR5700] - BSA and Azide free (ab178076)

This data was developed using the same antibody clone in a different buffer formulation (ab133619).

Lanes 1-2: Merged signal (red and green). Green - ab133619 observed at 37 kDa. Red - loading control ab8245 observed at 37 kDa.

 ab133619 Anti-Fas antibody [EPR5700] was shown to specifically react with Fas in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265260 (knockout cell lysate ab256911) was used. Wild-type and Fas knockout samples were subjected to SDS-PAGE. ab133619 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

Anti-Fas antibody [EPR5700] (ab133619)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling Fas with ab133619 at a dilution of 1:500. Heat mediated antigen retrieval was performed using AR9 antigen retrieval solution, and microwave treatment for 15 min at 20% power. Anti-Rabbit/Mouse HRP polymer (PerkinElmer Opal Polymer HRP Ms Plus Rb) was used as secondary antibody. Opal tyramide amplification was performed using Opal 520 fluorophore. Counterstained with DAPI stain. Image scanned with Vectra 3.0 and analyzed via Phenochart software.
This image was courteously provided by Dr. Houssein Abdul Sater, Georgia Cancer Center.

Immunohistochemical analysis of paraffin embedded Human tonsil tissue labelling CD95 with ab133619 antibody at a dilution of 1/250.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133619).

Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

Clone EPR5700 (ab178076) has been successfully conjugated by Abcam. This image was generated using Anti-Fas antibody [EPR5700] (Alexa Fluor® 488). Please refer to ab204670 for protocol details.

ab204670 staining Fas in Raji cells. The cells were fixed with 80% methanol (5 min) and then incubated in 1%BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated overnight at +4°C with ab204670 at 1/100 dilution (shown in green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 594), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

This product also gave a positive signal under the same testing conditions in Raji cells fixed with 4% formaldehyde (10 min).

Clone EPR5700 (ab178076) has been successfully conjugated by Abcam. This image was generated using Anti-Fas antibody [EPR5700] (Alexa Fluor® 647). Please refer to ab204671 for protocol details.

ab204671 staining Fas in Raji cells. The cells were fixed with 4% formaldehyde (10 min) and then incubated in 1%BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated overnight at +4°C with ab204671 at 1/50 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).