Immunofluorescent analysis of LIN28 (green) in Hela cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes at room temperature and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a LIN28 polyclonal antibody (Product # PA1-096 ) at a dilution of 1:100 and incubated overnight in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody for 45 minutes at room temperature in the dark. F-actin (red) was stained with a fluorescent phalloidin and nuclei (blue) were stained with DAPI. Images were taken at a 60X 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.
PA1-096 has been successfully used in Western blot, immunofluorescence, immunohistochemistry, flow cytometry, 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.