LIN-28A Antibody (14E6-4E6)

Flow Cytometry: LIN-28A Antibody (14E6-4E6) [NBP2-22481]

Flow Cytometry: LIN-28A Antibody (14E6-4E6) [NBP2-22481] - Analysis of Lin28 (blue histogram) on H9 embryonic stem cells. To generate single cells suspensions, colonies were treated with TrypLE cell dissociation enzyme for 5 minutes at 37C. Cells were incubated with a Lin28 monoclonal antibody 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 500ul of FACS buffer containing 10ul of 4% paraformaldehyde.

Chromatin Immunoprecipitation: LIN-28A Antibody (14E6-4E6) [NBP2-22481] - Analysis performed using cross-linked chromatin from rat hepatoma cells treated with insulin. IP performed using a multiplex microplate Matrix ChIP assay of LIN28 monoclonal antibody. Chromatin aliquots from cells were used per ChIP pull-down. Quantitative PCR data done in quadruplicate using 1ul of DNA in 2ul SYBR real-time PCR reactions containing primers to amplify -15kb upstream of Egr1 or exon-1 or exon-2-3 of Egr1. Quantitation of immunoprecipitated chromatin is presented as signal relative to the total amount of input chromatin. Results represent the mean +/- SEM. A schematic representation of the rat Egr-1 locus is shown; oxes represent exons (black boxes = translated, white boxes = untranslated), the zigzag line represents an intron, and the straight line represents upstream sequence. Regions amplified by Egr-1 primers are represented by black bars. Data courtesy of the Innovators Program.

Immunocytochemistry/ Immunofluorescence: LIN-28A Antibody (14E6-4E6) [NBP2-22481]

Immunocytochemistry/Immunofluorescence: LIN-28A Antibody (14E6-4E6) [NBP2-22481] - Analysis of Lin28 (green) in H9 embryonic stem 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 Lin28 monoclonal antibody at a dilution of 1:200 overnight at 4C, 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.

Immunocytochemistry/ Immunofluorescence: LIN-28A Antibody (14E6-4E6) [NBP2-22481]

Immunocytochemistry/Immunofluorescence: LIN-28A Antibody (14E6-4E6) [NBP2-22481] - Analysis of Lin28 (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 Lin28 monoclonal antibody at a dilution of 1:200 overnight at 4C, 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.

Immunohistochemistry-Paraffin: LIN-28A Antibody (14E6-4E6) [NBP2-22481]

Immunohistochemistry-Paraffin: LIN-28A Antibody (14E6-4E6) [NBP2-22481] - Analysis showing staining in the nucleus and cytoplasm of mouse testis tissue (right) compared with a negative control without primary antibody (left).

Flow Cytometry: LIN-28A Antibody (14E6-4E6) [NBP2-22481]

Flow Cytometry: LIN-28A Antibody (14E6-4E6) [NBP2-22481] - Analysis of Lin28 (blue histogram) on HEL 11.4 induced IPS cells. To generate single cells suspensions, colonies were treated with TrypLE cell dissociation enzyme for 5 minutes at 37C. Cells were incubated with a Lin28 monoclonal antibody 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 500ul of FACS buffer containing 10ul of 4% paraformaldehyde.

Immunoprecipitation: LIN-28A Antibody (14E6-4E6) [NBP2-22481]

Immunoprecipitation: LIN-28A Antibody (14E6-4E6) [NBP2-22481] - Analysis of LIN28 was performed. Antigen-antibody complexes were formed by incubating 700ug of lysate with 5 ug of an LIN28 monoclonal antibody overnight on a rocking platform at 4C. The immune complexes were captured on 50 ul Protein A/G Agarose was loaded as a positive control for detection. Samples were resolved on a 4-20% Tris-HCl polyacrylamide gel, transferred to a PVDF membrane, and blocked with 5% BSA/TBS-0.1%Tween for at least 1 hour. The membrane was probed with a LIN28 monoclonal antibody at a dilution of 1:1000 overnight rotating at 4C, washed in TBST, and probed with Clean-blot IP Detection Reagent at a dilution of 1:1000 for at least 1 hour.