Immunofluorescent analysis of Lin28 (red) 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 polyclonal antibody (Product # PA1-096) at a dilution of 1:200 overnight at 4°C, washed with PBST, and incubated with a fluorescently-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.
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Full-length human recombinant protein expressed in bacteria|
|Storage buffer||PBS with 1mg/ml BSA, 30% glycerol|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:100|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20-1:50|
|Immunoprecipitation (IP)||5 µg, 1:1000|
|Western Blot (WB)||1:1000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
PA1-096X has been successfully used in Western blot, immunofluorescence, IHC (P) , flow cytometery, and immunoprecipitation applications on human and mouse samples.
LIN28A/B are expressed in various cancers and ES cells. LIN28 is a cytoplasmic protein, that acts as a 'translational enhancer', driving specific mRNAs to polysomes and thus increasing the efficiency of protein synthesis. LIN28 is a negative regulator of microRNA (miRNA) biogenesis by specifically binding the precursor of let-7 (pre-let-7). Let-7 miRNAs have been implicated in repression of oncogenes such as ras and myc. LIN28 expression decreases during differentiation of ES cells or upon induction of neuronal differentiation by retinoic acid.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.