Western blot analysis of ovalbumin was performed by loading the indicated amounts of ovalbumin protein (Product # 77120), and 10ul of PageRuler Prestained Protein Ladder (Product # 26616) per well onto a Novex® 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a PVDF membrane (Product # 88518) using the G2 Blotter (Product # 62288), and blocked with 5% milk in TBST for at least 1 hour at room temperature. A doublet of Ovalbumin protein was detected between 40-50kD using an HRP-conjugated ovalbumin polyclonal antibody (Product # PA1-196-HRP) at a dilution of 1:5000 in blocking buffer. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080). NOTE: The presence of a doublet may be due to the presence of two different glycosylated forms of ovalbumin within the purified protein preparation.
|Tested species reactivity||Chicken|
|Host / Isotype||Rabbit / IgG|
|Storage buffer||PBS with proprietary stabilizer|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:1000-1:5000|
* 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 Western blot, PA1-196-HRP detects ovalbumin at ~45kD. Multiple bands may be observed, which are likely due to the presence of multiple glycosylated and/or aggregated species of ovalbumin within a particular protein preparation.
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 whether ovalbumin at this site plays a specific role in shell mineralization.
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