Dip-stick western blot

Patent metadata

Field	Value
Publication number	US-10107780-B2
Application number	US-201514644527-A
Country	US
Kind code	B2
Filing date	Mar 11, 2015
Priority date	Mar 11, 2014
Publication date	Oct 23, 2018
Grant date	Oct 23, 2018

Abstract

Methods, kits, and systems are provided for separating, immobilizing, and/or detecting analytes of one or more samples using dipsticks. A ‘dipstick’ is an object that can be embedded and subsequently removed from a separation medium, and to which analytes can be immobilized while the object is embedded in the separation medium. Examples of separation media include an electrophoresis gel of any format and a stationary phase for column chromatography. Embodiments of the present methods include applying a sample to a separation medium; separating analytes of the sample in the separation medium along a separation axis; immobilizing the analytes on a dipstick embedded in the separation medium; removing the dipstick from the separation medium; and detecting the analytes immobilized on the removed dipstick.

First claim

What is claimed is:

1. A method of separating and detecting analytes of a sample, the method comprising:
applying a sample to a separation medium, wherein the separation medium is a polyacrylamide gel;
separating analytes of the sample in the separation medium to generate a distribution of the analytes along a separation axis;
immobilizing the analytes on a dipstick embedded in part of the separation medium in which analytes become distributed such that the dipstick or an elongated portion thereof is aligned parallel to the separation axis, wherein distribution of analytes immobilized on the dipstick preserves the distribution of analytes as occurred in the separation medium, wherein the dipstick is embedded before the separating;
removing the dipstick from the separation medium; and
detecting the analytes immobilized on the removed dipstick,
wherein separating analytes of the sample comprises performing electrophoresis, electroosmosis, or isoelectric focusing.

2. The method of claim 1 , wherein the separation medium comprises a gel in a tube or cassette.

3. The method of claim 1 , wherein the separation medium comprises a denaturant.

4. The method of claim 1 , wherein immobilizing the analytes on the dipstick comprises covalently linking the analytes to the dipstick.

5. The method of claim 1 , wherein immobilizing the analytes on the dipstick comprises binding the analytes to an antibody.

6. The method of claim 1 , wherein immobilizing the analytes on the dipstick comprises depositing the analytes on a membrane coupled to the dipstick.

7. The method of claim 1 , wherein immobilizing the analytes on the dipstick comprises exposing the dipstick to fixation reagents.

8. The method of claim 1 , wherein:
the dipstick comprises an electrode, and immobilizing the analytes on the dipstick comprises energizing the electrode; or,
the dipstick comprises a magnet, and immobilizing the analytes on the dipstick comprises drawing magnetic labels linked to the analytes toward the magnet.

9. The method of claim 1 , wherein detecting the analytes immobilized on the removed dipstick comprises contacting the removed dipstick with a binding partner for one or more analytes of the sample, and detecting a signal indicative of binding between the binding partner and the one or more analytes.

10. The method of claim 1 , further comprising embedding the dipstick in the separation medium before separating the analytes.

11. The method of claim 1 , wherein the dipstick comprises a texture or beaded coating that increases the surface area of the dipstick.

12. The method of claim 1 , the analytes are proteins.

13. The method of claim 1 , wherein the separating is by charge or molecular weight of the analytes.

14. The method of claim 1 , wherein all of the analytes in the sample that are separated are immobilized on the dipstick.

15. The method of claim 1 , wherein the detecting comprises detecting the analytes while the analytes are immobilized on the removed dipstick.

16. The method of claim 1 , wherein the dipstick comprises a handle portion configured for mechanical manipulation of the dipstick.

Assignees

• Bio Rad Laboratories

Inventors

• Strong William

Classifications

• G01N27/44739 Primary

  Collecting the separated zones, e.g. blotting to a membrane or punching of gel spots · CPC title

• G01N30/90

  Plate chromatography, e.g. thin layer or paper chromatography · CPC title

• G01N33/54386

  Analytical elements · CPC title