- MA1-028 - Invitrogen Antibodies GATA3 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Gata3 (green) 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 Gata3 monoclonal antibody (Product # MA1-028) at a dilution of 1:100 (2 µg/mL) in 3% BSA solution (diluted in PBS) at 4C overnight, and then incubated with a secondary antibody (donkey anti mouse-AlexaFluor 488, green). 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 GATA3 (green) in MCF7 cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature. Cells were blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed without (left panel) or with (right panel) a GATA3 monoclonal antibody (Product # MA1-028) at a dilution of 1:50 for at least 1 hour at room temperature, washed with PBS, and incubated with a DyLight 488-conjugated goat anti-mouse IgG secondary antibody (Product # 35503) at a dilution of 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with DyLight-554 Phalloidin (Product # 21834) and nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of GATA3 (green) in SH-SY5Y cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature. Cells were blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed without (left panel) or with (right panel) a GATA3 monoclonal antibody (Product # MA1-028) at a dilution of 1:50 for at least 1 hour at room temperature, washed with PBS, and incubated with a DyLight 488-conjugated goat anti-mouse IgG secondary antibody (Product # 35503) at a dilution of 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with DyLight-554 Phalloidin (Product # 21834) and nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of GATA3 was performed using 70% confluent log phase MCF7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with GATA3 Mouse Monoclonal Antibody (1A12-1D9) (Product # MA1-028) at 5 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Chromatin Immunoprecipitation (ChIP) assay of endogenous GATA3 protein using Anti-GATA3 Antibody: ChIP was performed using Anti-GATA3 Monoclonal Antibody (1A12-1D9) (Product # MA1-028) 5 µg, on sheared chromatin from HCT116 cells using the MAGnify ChIP System kit (Product # 49-2024). Normal Mouse IgG was used as a negative IP control. The purified DNA was analyzed by qPCR using primers binding to promoter of HMBS and SAT2 satellite repeats. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunoprecipitation of GATA3 was performed on SH-SY5Y cells. Antigen-antibody complexes were formed by incubating 250 µg of SH-SY5Y whole cell lysate with 5 µg of a GATA3 monoclonal antibody (Product # MA1-028) overnight on a rocking platform at 4øC. The immune complexes were captured on 50 µL Protein A/G Agarose (Product # 20421), washed extensively, and eluted with 5X Lane Marker Reducing Sample Buffer (Product # 39000). Eluted sample and 25 µg of SH-SY5Y whole cell lysate (loading control) were resolved on a 4-20% Tris-HCl polyacrylamide gel, transferred to a PVDF membrane, and blocked with StartingBlock T20 (Product # 37543) for at least 1 hour. The membrane was probed with a GATA3 monoclonal antibody (Product # MA1-028) at a dilution of 1:1000 overnight rotating at 4øC, washed in TBST, and probed with an HRP- conjugated, light chain specific goat anti-mouse IgG at a dilution of 1:20,000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080). Images were acquired on a Thermo Scientific myECL Imager (Product # 62236).

MA1-028

Antibody data