Multifunctional protein molecular weight ladders

Patent metadata

Field	Value
Publication number	US-9663554-B2
Application number	US-201514857661-A
Country	US
Kind code	B2
Filing date	Sep 17, 2015
Priority date	Sep 19, 2014
Publication date	May 30, 2017
Grant date	May 30, 2017

Abstract

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.

First claim

What is claimed is:

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:

Assignees

• Li Cor Inc

Inventors

• Padhye Nisha
• Skopp Rose
• Wang Ying

Classifications

• C07K7/06 Primary

  having 5 to 11 amino acids · CPC title

• C12Q1/28

  involving peroxidase · CPC title

• G01N33/582

  with fluorescent label · CPC title

• C07K14/795

  Porphyrin- or corrin-ring-containing peptides · CPC title

• G01N2550/00

  Electrophoretic profiling, e.g. for proteome analysis · CPC title