Western blot analysis of ovalbumin was performed by loading the indicated amounts of recombinant ovalbumin protein (Product # 77126) per well, and 10ul of PageRuler Plus Prestained Protein Ladder (Product # 26619) onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane using the G2 Fast Blotter (Product # 62288), and blocked with StartingBlock T20 (TBS) Blocking Buffer (Product # 37543) for 1 hour at room temperature. Ovalbumin was detected at ~45kDa after probing with an ovalbumin polyclonal antibody (Product # PA1-196) at a dilution of 1:1000 in StartingBlock T20 (TBS) Blocking Buffer (Product # 37543) overnight at 4C on a rocking platform, washing in TBST, and probing with an HRP-conjugated goat anti-rabbit IgG secondary antibody (Product # 31460) at a dilution of 1:40,000 for 1 hour. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080). NOTE: In addition to detecting ovalbumin monomer, this antibody also detects aggregated ovalbumin proteins at higher molecular weights.
|Tested species reactivity||Chicken|
|Host / Isotype||Rabbit / IgG|
|Storage buffer||PBS with 1mg/ml BSA, 30% glycerol|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Western Blot (WB)||1:100-1:1000|
* 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.
By WB, PA1-196 detects ovalbumin at ~45kD.
Chicken ovalbumin is the major protein in the "white" of the egg (and a favorite antigen in immunological research). Egg white contains a variety of proteins including ovalbumin, conalbumin, ovomucoi and lysozyme. It belonds to the serpin family and the Ov serpin subfamily. Ovalbumin can cause an allergic reaction in humans. Ovalbumin has been implicated in the development of the egg shell. Immunohistochemistry revealed that ovalbumin is found only in the mammillary bodies of decalcified shell, and is not distributed throughout the shell matrix. These results indicate that ovalbumin is present during the initial phase of shell formation and becomes incorporated into the protein matrix of the mammillary bodies. However, it is not yet clear wheather ovalbumin at this site plays a specific role in shell mineralisation.
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