Multifunctional protein molecular weight ladders
Multifunctional molecular weight protein ladders and methods of making thereof are disclosed herein that are useful for determining the molecular weight of a test protein and/or the relative mass or amount of the test protein in a protein separation assay, such as gel electrophoresis or western blotting. Also included are compounds of Formula I (e.g., mono acetylated MP-11 NHS ester) that may be used to label purified proteins of the protein ladder. The MP-11 label protein ladder can be detected on a blotting membrane by exposing the microperoxidase to a suitable substrate, such as a chromogenic substrate or a chemiluminescent substrate.
1. A protein standard, said protein standard comprising a plurality of purified proteins having differing molecular weights; and
wherein one or more proteins from the plurality of purified proteins is covalently linked to a mono microperoxidase N-hydroxy succinimidyl (NHS) ester of the formula:
wherein n is 0-10, and the covalent linkage is between an amine from the one or more proteins and the N-hydroxy succinimidyl (NHS) ester group of the mono microperoxidase N-hydroxy succinimidyl (NHS) ester.
2. The protein standard of claim 1 , wherein the protein standard comprises at least 5 different purified proteins.
3. The protein standard of claim 1 , wherein the protein standard comprises at least 10 different purified proteins.
4. The protein standard of claim 1 , wherein the range of molecular weights of the protein standard is about 5 kDa to about 280 kDa.
5. The protein standard of claim 1 , wherein the range of molecular weights of the protein standard is about 8 kDa to about 260 kDa.
6. The protein standard of claim 1 , wherein at least 2 of the purified proteins of the protein standard are covalently linked to the mono microperoxidase NHS ester.
7. The protein standard of claim 1 , wherein at least 3 of the purified proteins of the protein standard are covalently linked to the mono microperoxidase NHS ester.
8. The protein standard of claim 1 , wherein at least 4 of the purified proteins of the protein standard are covalently linked to the mono microperoxidase NHS ester.
9. The protein standard of claim 1 , wherein at least 5 of the purified proteins of the protein standard are covalently linked to the mono microperoxidase NHS ester.
10. The protein standard of claim 1 , wherein at least 10 of the purified proteins of the protein standard are covalently linked to the mono microperoxidase NHS ester.
11. The protein standard of claim 1 , wherein the mono microperoxidase NHS ester is a mono MP-11 NHS ester of formula Ia: