Western blot analysis of WDR76 was performed by loading 500 ng of whole cell extract from equal numbers of wild type mouse embryonic E14 stem cells (E14 mESCs) and E14 mESCs in which the WDR76 locus has been truncated via CRISPR editing. Sample was loaded in reducing sample buffer (5x = 10% SDS, 50% glycerol (v/v), 0.25% bromophenol blue (w/v) and 10 mM beta mercaptoethanol) on a 14% polyacrylamide gel. Proteins were transferred 0.22 micron PVDF membrane in Towbins buffer (25 mM Tris, 192 mM glycine, 20% methanol (v/v), 0.1% SDS (v/v)) at 100 mA for 30 minutes. WDR76 was detected at approximately 70 kDa using a WDR76 polyclonal antibody (Product # PA5-24147) at a dilution of 1:1,000. This incubation proceeded overnight and was followed by incubation with a 1:10,000 dilution of a secondary HRP-conjµgated goat anti mouse IgG antibody (Product# 31432) at a dilution of 1:20,000. Chemiluminescent detection was performed. The loading level of the wildtype and CRISPR targeted E14 mESCs was compared by blotting for histone H3 using a 1:20,000 dilution of this antibody and the same HRP-conjµgated goat anti mouse IgG antibody as described above. The WDR76 confirmed atthe protein level that while WDR76 is detected in the wildtype E14 mESCs, the CRISPR targeted line does not contain fµll length WDR76. Data provided courtesy of Kate Malecek and Alex Ruthenburg..
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||KLH conjugated synthetic peptide between 455-484 amino acids from the C-terminal region of human WDR76|
|Purification||Antigen affinity chromatography|
|Contains||0.09% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|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.
PA5-24147 detects WDR76 in WB application.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.