Rabbit YWHAE Polyclonal Antibody | anti-YWHAE antibody
Anti-YWHAE Antibody
IHC-P: 2-5ug/ml|Mouse, Rat|
ICC/IF: 5ug/ml|Human|
FC/FACS/FCM: 1-3ug/1x106 cells|Human|
Direct ELISA: 0.1-0.5ug/ml|Human|
Western Blot (WB)
(Figure 1. Western blot analysis of YWHAE using anti-YWHAE antibody (MBS1753454).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: human Hela whole cell lysatesLane 2: human Jurkat whole cell lysatesLane 3: human HepG2 whole cell lysatesLane 4: human SH-SY5Y whole cell lysatesLane 5: human HEK293 whole cell lysatesLane 6: human SW620 whole cell lysatesLane 7: human A549 whole cell lysatesLane 8: human Raji whole cell lysatesLane 9: rat brain tissue lysatesLane 10: mouse brain tissue lysatesLane 11: mouse NIH/3T3 whole cell lysatesLane 12: mouse RAW264. 7 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti-YWHAE antigen affinity purified polyclonal antibody (Catalog # MBS1753454) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for YWHAE at approximately 28KD. The expected band size for YWHAE is at 28KD. )
Immunohistochemistry (IHC)
(Figure 2. IHC analysis of YWHAE using anti YWHAE antibody (MBS1753454).YWHAE was detected in paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-YWHAE Antibody (MBS1753454) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )
Flow Cytometry (FC/FACS)
(Figure 3. Flow Cytometry analysis of A549 cells using anti-YWHAE antibody (MBS1753454).Overlay histogram showing A549 cells stained with MBS1753454 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-YWHAE Antibody (MBS1753454,1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Immunofluorescence (IF)
(Figure 4. IF analysis of YWHAE using anti- YWHAE antibody (MBS1753454).YWHAE was detected in immunocytochemical section of MCF-7 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti- YWHAE Antibody (MBS1753454) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )
2. Cardoso, C., Leventer, R. J., Ward, H. L., Toyo-oka, K., Chung, J., Gross, A., Martin, C. L., Allanson, J., Pilz, D. T., Olney, A. H., Mutchinick, O. M., Hirotsune, S., Wynshaw-Boris, A., Dobyns, W. B., Ledbetter, D. H. Refinement of a 400-kb critical region allows genotypic differentiation between isolated lissencephaly, Miller-Dieker syndrome, and other phenotypes secondary to deletions of 17p13. 3. Am. J. Hum. Genet. 72: 918-930, 2003.
3. Chong, S. S., Tanigami, A., Roschke, A. V., Ledbetter, D. H. 14-3-3-epsilon has no homology to LIS1 and lies telomeric to it on chromosome 17p13. 3 outside the Miller-Dieker syndrome chromosome region. Genome Res. 6: 735-741, 1996.
NCBI and Uniprot Product Information
NCBI Description
This gene product belongs to the 14-3-3 family of proteins which mediate signal transduction by binding to phosphoserine-containing proteins. This highly conserved protein family is found in both plants and mammals, and this protein is 100% identical to the mouse ortholog. It interacts with CDC25 phosphatases, RAF1 and IRS1 proteins, suggesting its role in diverse biochemical activities related to signal transduction, such as cell division and regulation of insulin sensitivity. It has also been implicated in the pathogenesis of small cell lung cancer. Two transcript variants, one protein-coding and the other non-protein-coding, have been found for this gene. [provided by RefSeq, Aug 2008]
Uniprot Description
14-3-3 epsilon: a protein of the 14-3-3 family of proteins which mediate signal transduction by binding to phosphoserine-containing proteins. A multifunctional regulator of the cell signaling processes.
Protein type: Adaptor/scaffold
Chromosomal Location of Human Ortholog: 17p13.3
Cellular Component: kinesin complex; cytoplasmic vesicle membrane; focal adhesion; mitochondrion; membrane; axon; melanosome; cytosol
Molecular Function: protein domain specific binding; protein binding; enzyme binding; potassium channel regulator activity; protein heterodimerization activity; histone deacetylase binding; phosphoprotein binding; phosphoserine binding
Biological Process: substantia nigra development; nerve growth factor receptor signaling pathway; viral reproduction; apoptosis; organelle organization and biogenesis; hippocampus development; neuron migration; mitotic cell cycle; cerebral cortex development; regulation of caspase activity; G2/M transition of mitotic cell cycle; regulation of the rate of heart contraction by hormone; protein targeting
Disease: Miller-dieker Lissencephaly Syndrome
