Use of protein G for preparation and characterization of rabbit antibodies against rat adipose tissue hormone-sensitive lipase
Fredrikson, G ; Nilsson, S LU ; Olsson, H LU
; Björck, L
LU
; Akerström, B
LU
and Belfrage, P
LU
(1987)
In Journal of Immunological Methods
97(1).
p.65-70
- Abstract
The newly described immunoglobulin G-binding streptococcal surface protein, protein G, was used to prepare and characterize rabbit antibodies. The antibodies were directed against rat hormone-sensitive lipase, the rate-limiting enzyme in the hydrolysis of the triacylglycerols stored in adipose tissue. Antiserum was obtained after two injections with 20 micrograms enzyme protein, and the immunoglobulin fraction was obtained using a protein G-based solid-phase radioimmunoassay. The hydrolysis of acylglycerols by the enzyme was inhibited by the antibodies, and the enzyme could be efficiently removed from a solution using the antibodies and heat-killed streptococci expressing surface protein G. By Western blot and detection with... (More)
The newly described immunoglobulin G-binding streptococcal surface protein, protein G, was used to prepare and characterize rabbit antibodies. The antibodies were directed against rat hormone-sensitive lipase, the rate-limiting enzyme in the hydrolysis of the triacylglycerols stored in adipose tissue. Antiserum was obtained after two injections with 20 micrograms enzyme protein, and the immunoglobulin fraction was obtained using a protein G-based solid-phase radioimmunoassay. The hydrolysis of acylglycerols by the enzyme was inhibited by the antibodies, and the enzyme could be efficiently removed from a solution using the antibodies and heat-killed streptococci expressing surface protein G. By Western blot and detection with 125I-protein G, the antibodies were found to selectively bind to hormone-sensitive lipase and to a smaller extent to two minor contaminants, possibly proteolytic fragments of the lipase. The amount of 125I-labelled protein G bound to the lipase on the blot was quantitatively related to the amount of enzyme protein down to the detection limit 10 ng.
(Less)
- author
- Fredrikson, G
; Nilsson, S
LU
; Olsson, H
LU
; Björck, L
LU
; Akerström, B
LU
and Belfrage, P
LU
- organization
- publishing date
- 1987-02-26
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Adipose Tissue/enzymology, Animals, Antibodies/analysis, Antigen-Antibody Complex, Bacterial Proteins/immunology, Hormones/pharmacology, Immunoglobulin G/immunology, Rabbits/immunology, Radioimmunoassay/methods, Rats, Sterol Esterase/analysis
- in
- Journal of Immunological Methods
- volume
- 97
- issue
- 1
- pages
- 65 - 70
- publisher
- Elsevier
- external identifiers
-
- pmid:3819438
- scopus:0023124216
- ISSN
- 0022-1759
- DOI
- 10.1016/0022-1759(87)90106-2
- language
- English
- LU publication?
- yes
- id
- 5c140892-f243-43d3-a3a0-ba60f6ece816
- date added to LUP
- 2019-05-22 10:32:12
- date last changed
- 2024-01-01 06:54:41
@article{5c140892-f243-43d3-a3a0-ba60f6ece816,
abstract = {{<p>The newly described immunoglobulin G-binding streptococcal surface protein, protein G, was used to prepare and characterize rabbit antibodies. The antibodies were directed against rat hormone-sensitive lipase, the rate-limiting enzyme in the hydrolysis of the triacylglycerols stored in adipose tissue. Antiserum was obtained after two injections with 20 micrograms enzyme protein, and the immunoglobulin fraction was obtained using a protein G-based solid-phase radioimmunoassay. The hydrolysis of acylglycerols by the enzyme was inhibited by the antibodies, and the enzyme could be efficiently removed from a solution using the antibodies and heat-killed streptococci expressing surface protein G. By Western blot and detection with 125I-protein G, the antibodies were found to selectively bind to hormone-sensitive lipase and to a smaller extent to two minor contaminants, possibly proteolytic fragments of the lipase. The amount of 125I-labelled protein G bound to the lipase on the blot was quantitatively related to the amount of enzyme protein down to the detection limit 10 ng.</p>}},
author = {{Fredrikson, G and Nilsson, S and Olsson, H and Björck, L and Akerström, B and Belfrage, P}},
issn = {{0022-1759}},
keywords = {{Adipose Tissue/enzymology; Animals; Antibodies/analysis; Antigen-Antibody Complex; Bacterial Proteins/immunology; Hormones/pharmacology; Immunoglobulin G/immunology; Rabbits/immunology; Radioimmunoassay/methods; Rats; Sterol Esterase/analysis}},
language = {{eng}},
month = {{02}},
number = {{1}},
pages = {{65--70}},
publisher = {{Elsevier}},
series = {{Journal of Immunological Methods}},
title = {{Use of protein G for preparation and characterization of rabbit antibodies against rat adipose tissue hormone-sensitive lipase}},
url = {{http://dx.doi.org/10.1016/0022-1759(87)90106-2}},
doi = {{10.1016/0022-1759(87)90106-2}},
volume = {{97}},
year = {{1987}},
}