Key facts
Start here
Agilent Analysis of Hsp47 a Collagen Chaperone by Size Exclusion Chromatography (SEC) Manual
This document details the analysis of Heat shock protein 47 (Hsp47), a collagen-specific molecular chaperone, using Size Exclusion Chromatography (SEC) with UV detection. The study focuses on identifying different species present in a native mouse Hsp47 sample, including monomers, aggregates, and degraded forms.
Read onlineAgilent ProSec 3008 specifications
| Column | Agilent ProSec 3008, 300 x 7.5 mm |
|---|---|
| Part Number | PL1147-6501 |
| Flow Rate | 1 mL/min |
| Temperature | 5°C |
| Detection | UV at 280 nm |
| Concentration | 2mg/ml |
| Eluent | 0.1 M KH2PO4 containing 0.3 M NaCl at pH 7.0 |
Source: Agilent Analysis of Hsp47 a Collagen Chaperone by Size Exclusion Chromatography (SEC) Manual
All Agilent ProSec 3008 documents by type
How to use the product for one specific task, with worked examples.
Questions answered in these manuals
What is the molecular weight of the Hsp47 monomer?
The monomer of Hsp47 has a molecular weight of 47 kDa.
From: Agilent Analysis of Hsp47 a Collagen Chaperone by Size Exclusion Chromatography (SEC) Manual
What does the chromatogram indicate about the native Hsp47 sample?
The sample exhibited multiple signals indicating various oligomeric states, including aggregation and protein degradation.
From: Agilent Analysis of Hsp47 a Collagen Chaperone by Size Exclusion Chromatography (SEC) Manual
What denaturants are used in this analysis?
The analysis uses urea at concentrations of 2 M introduced for two hours and 8 M introduced overnight.
From: Agilent ProSEC 3008 Application Note: Progressive Denaturation of Globular Proteins in Urea
What effect does urea have on globular proteins?
Urea instigates protein unfolding, producing higher molecular weight species due to the proteins adopting less compact structures and aggregation through disulfide bond formation.
From: Agilent ProSEC 3008 Application Note: Progressive Denaturation of Globular Proteins in Urea