Western blot analysis of Lin28 was performed by loading 50ug of NCCIT and HeLa cell lysates per well onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane. The membrane was blocked, and probed with an HRP-conjugated Lin28 monoclonal antibody (Product # MA1-016-HRP) at a dilution of 1:1000 overnight at 4C. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34075).
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Mouse / IgG|
|Immunogen||Full-length human recombinant protein expressed in bacteria|
|Storage buffer||proprietary buffer with proprietary stabilizer|
|Storage Conditions||4° C, do not freeze|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:250-1:1000|
* 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-HRP has been successfully used in Western blot 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.
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