Aug 15, 2020
Protein isolation from the extract of coelomic fluid of the Lumbricus terrestris.
- 1University of the West Indies St. Augustine
- University of the West Indies
- angel.vaillant@sta.uwi.edu
Protocol Citation: Angel A Justiz-Vaillant 2020. Protein isolation from the extract of coelomic fluid of the Lumbricus terrestris.. protocols.io https://dx.doi.org/10.17504/protocols.io.bjsaknae
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: August 15, 2020
Last Modified: August 15, 2020
Protocol Integer ID: 40482
Six earthworms, Lumbricus terrestris, after removal from the earth are placed in a 15 ml glass tube and washed thoroughly with PBS, pH 7.4.
The body of the annelid is pierced with an 18-gauge needle.
The coelomic fluid content of the annelid is added to 2 ml of PBS, pH 7.4.
After incubation at 4oC for 1h, 0.9 ml chloroform is added.
The mixture is centrifuged at 1000 x g for 5 min at 4oC.
The supernatant (1.1 ml) is collected and equal volume of cold ethanol is added drop wise to the preparation.
The mixture is then incubated at 4oC for 12 h.
Then, it is centrifuged at 4oC for 5 min.
The pellet (protein extract) is resuspended in 0.3 ml of PBS, pH 7.4.
The preparation is dialyzed against 1L of PBS, pH 7.4 for 12 h at 4oC.
The protein concentration is assessed by the Bradford method or any available method.