- 720030 - Invitrogen Antibodies SALL4 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Sall4 (red) in human ESCs WIRB3 and HES2. The cells were fixed with 300 uL 4% paraformaldehyde solution for 15 min at RT in the dark, permeabilized with 300 uL 0.25% TritonX-100 solution (diluted in PBS) for 5 min at RT, and blocked with 300 uL 10% BSA solution (diluted in PBS) for 30 min in a 37C incubator. Cells were stained with a Sall4 polyclonal antibody (Product # 720030) at a dilution of 1:250 (2 µg/mL) in 3% BSA solution (diluted in PBS) at 4C overnight, and then incubated with a secondary antibody (Red). Nuclei (blue) was stained with DAPI (Product # D1306). Images were taken on a Leica DM5500 microscope at 10X magnification. Data courtesy of Dr. Jianlong Wang's lab.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence was performed on fixed and permeabilized A-431cells for detection of Sall4 using Anti-Sall4 Rabbit Polyclonal Antibody (Product # 720030, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 0.4 µg/mL, 1:2500). Panel a) shows representative cells that were stained for detection and localization of Sall4 protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938, 1:50). Panel c) represents cytoskeletal F-actin staining using Alexa Fluor® 594 Phalloidin (Product # A12381, 1:200). Panel d) is a composite image of Panels a, b and c clearly demonstrating nuclear localization of Sall4. Panel e) represents control cells with no primary antibody to assess background.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Sall4 (green) in Episomal Human iPSC cells. The cells were fixed with formalin for 15 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and blocked with 3% BSA in PBS (Product # 37525) for 30 minutes at room temperature. Cells were stained with a Sall4 polyclonal antibody (Product # 720030), at a dilution of 2 µg/mL for one hour at room temperature, and then incubated with a goat anti-rabbit Superclonal IgG secondary antibody, Alexa Fluor 488 conjugate (Product # A27034) at a dilution of 1:1000 for 30 minutes at room temperature (green). Nuclei (blue) were counterstained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific EVOS FL at 40X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Sall4 (red) in human ESCs WIRB3 and HES2. The cells were fixed with 300 uL 4% paraformaldehyde solution for 15 min at RT in the dark, permeabilized with 300 uL 0.25% TritonX-100 solution (diluted in PBS) for 5 min at RT, and blocked with 300 uL 10% BSA solution (diluted in PBS) for 30 min in a 37C incubator. Cells were stained with a Sall4 polyclonal antibody (Product # 720030) at a dilution of 1:250 (2 µg/mL) in 3% BSA solution (diluted in PBS) at 4C overnight, and then incubated with a secondary antibody (Red). Nuclei (blue) was stained with DAPI (Product # D1306). Images were taken on a Leica DM5500 microscope at 10X magnification. Data courtesy of Dr. Jianlong Wang's lab.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Sall4 (green) in Episomal Human iPSC cells. The cells were fixed with formalin for 15 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and blocked with 3% BSA in PBS (Product # 37525) for 30 minutes at room temperature. Cells were stained with a Sall4 polyclonal antibody (Product # 720030), at a dilution of 2 µg/mL for one hour at room temperature, and then incubated with a goat anti-rabbit Superclonal IgG secondary antibody, Alexa Fluor 488 conjugate (Product # A27034) at a dilution of 1:1000 for 30 minutes at room temperature (green). Nuclei (blue) were counterstained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific EVOS FL at 40X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence was performed on fixed and permeabilized A-431cells for detection of Sall4 using Anti-Sall4 Rabbit Polyclonal Antibody (Product # 720030, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (Heavy Chain) Superclonal Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 0.4 µg/mL, 1:2500). Panel a) shows representative cells that were stained for detection and localization of Sall4 protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938, 1:50). Panel c) represents cytoskeletal F-actin staining using Alexa Fluor® 594 Phalloidin (Product # A12381, 1:200). Panel d) is a composite image of Panels a, b and c clearly demonstrating nuclear localization of Sall4. Panel e) represents control cells with no primary antibody to assess background.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Sall4 showing staining in the nucleus of paraffin-embedded mouse testis tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Sall4 Rabbit Polyclonal Antibody (Product # 720030) diluted in 3% BSA-PBS at a dilution of 1:100 for 1 hour at 37ºC in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

720030