Western blot analysis of c-Ret was performed using membrane enriched extracts (30 ug lysate) of SH-SY5Y (Lane 1), SK-N-AS (Lane 2) and Neuro-2a (Lane 3). The blots were probed with Anti-c-Ret Mouse Monoclonal Antibody (Product # AHO1482, 2 µg/ml) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.4 µg/ml, 1:2500 dilution). A 170 kDa band corresponding to c-Ret was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0302BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with overnight wet transfer system. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Mouse / IgG1, kappa|
|Immunogen||Recombinant fragment of human c-Ret extracellular domain expressed in E. coli.|
|Storage buffer||PBS, pH 7.2, with 1% BSA|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|Western Blot (WB)||1-3 µg/ml|
* 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.
RET, a member of the cadherin superfamily, is one of the receptor tyrosine kinases, which are cell-surface molecules that transduce signals for cell growth and differentiation. This protein plays a crucial role in neural crest development, and the gene can undergo oncogenic activation in vivo and in vitro by cytogenetic rearrangement. Mutations i are associated with the disorders multiple endocrine neoplasia, type IIA, multiple endocrine neoplasia, type IIB, Hirschsprung disease, and medullary thyroid carcinoma.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.