Transcription-Specific Antibodies

Transcription factors are proteins that bind to specific DNA sequences and regulate the transcription of mRNA from DNA by activating or inhibiting RNA polymerase. Transcription factors contain more than one DNA-binding domain, allowing them to bind to specific DNA sequences near the genes that they regulate. Invitrogen transcription-specific antibodies are designed to dependably detect key transcription factor targets. Each antibody is validated for use in various applications.

Key transcription-specific targets include:

Applications

Invitrogen transcription-specific antibodies are designed to dependably detect key transcription factor targets in various applications.

Immunofluorescent analysis of c-Myc (green) in HEL 11.4 induced IPS cells grown for a few days on Matrigel-coated chamber slides. Cells fixed in 4% paraformaldehyde were permeabilized with 0.1% Triton X-100 for 15 minutes at room temperature. Cells were probed with a c-Myc monoclonal antibody (Cat. No. MA1-980) at a dilution of 1:200 overnight at 4°C, washed with PBST, and incubated with a fluorescein-conjugated secondary antibody at a dilution of 1:100 for 1 hour at room temperature. Nuclei (blue) were stained with DAPI and cells were analyzed by fluorescence microscopy at 20X magnification.

Flow cytometric analysis of c-Myc (blue histogram) on H9 embryonic stem cells. To generate single cells suspensions, colonies were treated with TrypLE cell dissociation enzyme for 5 minutes at 37°C. Cells were incubated with a c-Myc monoclonal antibody (Cat. No. MA1-980) or mouse IgG (green histogram) at a dilution of 1:100 for 1 hour on ice, washed with PBS + 5% fetal calf serum (FACS buffer), and incubated with a fluorescein-conjugated secondary antibody at a dilution of 1:200 for 30 minutes on ice. Cells were washed with cold FACS buffer, resuspended in 500 µL of FACS buffer containing 10 µL of 4% paraformaldehyde, and analyzed on a flow cytometer.

Immunohistochemistry analysis of ETS (PS282/PS285) showing staining in the nucleus of paraffin-embedded human T cell lymphoma (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10 mM sodium citrate (pH 6.0), microwaved for 8-15 minutes. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 minutes at room temperature, washed with ddH2O and PBS, and then probed with a Phospho-ETS (Ser282, Ser285) Polyclonal Antibody (Cat. No. 44-1111G) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4°C 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.

Key transcription-specific targets include:

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