| Title |
Cloning, expression and purification of functionally active human angiopoietin-like protein 2
|
|---|---|
| Published in |
SpringerPlus, July 2014
|
| DOI | 10.1186/2193-1801-3-337 |
| Pubmed ID | |
| Authors |
Nada Farhat, Aida M Mamarbachi, Eric Thorin, Bruce G Allen |
| Abstract |
Angiopoietin-like protein 2 (Angptl2) is a secreted glycoprotein that has been implicated in angiogenesis, inflammation and atherosclerosis as well as enhancing the survival of human hematopoietic stem cells. Glycosylation of Angptl2 is required for biological activity and studies of angiopoietin-like protein 2 have been hindered by the lack of a source for the mature form of this protein. We describe a system that permits purification of the glycosylated form of human Angptl2 from conditioned media of stably transfected HEK 293 cells. To facilitate purification while retaining the integrity of Angptl2's endogenous N-terminal secretion signal peptide, GST was fused downstream of the Angptl2 coding sequence. Secreted Angptl2-GST was purified using a one-step glutathione-affinity purification scheme. The purity and identity of the resulting protein were confirmed by SDS-PAGE, immunoblotting, and mass spectrometry. N-Glycosidase treatment reduced the apparent molecular mass of Angptl2-GST on SDS-PAGE, confirming its glycosylation state. Purified human Angptl2-GST stimulated both HUVEC migration and microtubule formation in vitro. The yield of Angptl2-GST obtained was in quantities suitable for multiple applications including functional in vitro and in vivo assays. |
X Demographics
The data shown below were collected from the profile of 1 X user who shared this research output. Click here to find out more about how the information was compiled.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 1 | 100% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Scientists | 1 | 100% |
Mendeley readers
The data shown below were compiled from readership statistics for 16 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Montenegro | 1 | 6% |
| Unknown | 15 | 94% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 4 | 25% |
| Student > Bachelor | 3 | 19% |
| Student > Ph. D. Student | 3 | 19% |
| Other | 1 | 6% |
| Student > Doctoral Student | 1 | 6% |
| Other | 2 | 13% |
| Unknown | 2 | 13% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 8 | 50% |
| Agricultural and Biological Sciences | 3 | 19% |
| Immunology and Microbiology | 2 | 13% |
| Chemistry | 1 | 6% |
| Unknown | 2 | 13% |
Attention Score in Context
This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 02 August 2014.
All research outputs
#15,303,385
of 22,759,618 outputs
Outputs from SpringerPlus
#931
of 1,852 outputs
Outputs of similar age
#133,083
of 227,667 outputs
Outputs of similar age from SpringerPlus
#51
of 80 outputs
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