Immunofluorescent analysis of LIN28 in NCCIT and HeLa 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 with a LIN28 monoclonal antibody (Product # MA1-016) 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 # 35502). 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 ArrayScan at 20X magnification.
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Mouse / IgG|
|Immunogen||Full-length human recombinant protein expressed in bacteria|
|Storage buffer||PBS with 30% glycerol, 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||1-3 ul|
|Flow Cytometry (Flow)||1:100|
|Immunofluorescence (IF)||1:50 - 1:200|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20-1:200|
|Immunoprecipitation (IP)||5 µg|
|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.
MA1-016 has been successfully used in Western blot, immunofluorescence, ChIP, flow cytometry, IHC (P) 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.