Difference: TwodxClaudin15 (1 vs. 4)

Revision 403 Apr 2014 - Main.NicolasCoudray

Changed:
<
<
META TOPICPARENT name="2DCrystallizationExp"
>
>
META TOPICPARENT name="TwoDCrystallizationGroup"
 

2dx - Claudin-15: 2D crystallization

General 2dx information

  • the protein can be frozen (still leads to 3D crystals)
  • stable at 4C for 2 weeks at least
  • 23-25 kda
  • Detergents: they all behave similarly in the maltosides (DDM, UDM, DM) and fairly well in CHAPS. they can be exchanged into glucosides too. (The protein looks around 67kD off SEC in DM. Alex has noticed that when he prep the protein in OG, he gets some elution at 67kD, but the major peak in approx 350kD)

Properties:

  • sequence:
MSVAVETFGFFMSALGLLMLGLTLSNSYWRVSTVHGNVITTNTIFENLWYSCATDSLGVSNCWDFPSMLALSGYVQGCRALMITAILLGFLGLFLGMVGLRCTNVGNMDLSKKAKLLAIAGTLHILAGACGMVAISWYAVNITTDFFNPLYAGTKYELGPALYLGWSASLLSILGGICVFSTCCCSSKEEPATRAGLPYKPSTVVIPRATSDESDISFGKYGKNAYVGLVPR

X256 - Dec 6 2013 - Sparse Matrix

Protein in 20mM Tris pH 7.2, 150mM NaCl, 2% glycerol, 0.1% DM: Protein_acquisition_form-cldn15-Dec2013.doc.

Conditions: Sparse matrix + best conditions of CDN3 (3Dx crystals, cf CDN3 twikipage).

Details: X256-CDN15_Sparse.xlsx.

X256, results

Image selection: X256-CDN15_ImageSelection.pptx.

Mostly aggregates (E.coli worse), and small vesicles. 1 striated vesicles (but DOPC lipid, so likely a lipid crystal). Unclear if the protein is even reconstituted in the lipids.

  • Set ALLOWTOPICVIEW =

-- NicolasCoudray - 10 Dec 2013

Added:
>
>
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1386682829" name="X256-CDN15_Sparse.xlsx" path="X256-CDN15_Sparse.xlsx" size="587118" user="Main.NicolasCoudray" version="1"
 
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1386682821" name="Protein_acquisition_form-cldn15-Dec2013.doc" path="Protein_acquisition_form-cldn15-Dec2013.doc" size="60928" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1387304563" name="X256-CDN15_ImageSelection.pptx" path="X256-CDN15_ImageSelection.pptx" size="3874740" user="Main.NicolasCoudray" version="1"
Deleted:
<
<
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1386682829" name="X256-CDN15_Sparse.xlsx" path="X256-CDN15_Sparse.xlsx" size="587118" user="Main.NicolasCoudray" version="1"
 

Revision 317 Dec 2013 - Main.NicolasCoudray

 
META TOPICPARENT name="2DCrystallizationExp"

2dx - Claudin-15: 2D crystallization

General 2dx information

  • the protein can be frozen (still leads to 3D crystals)
  • stable at 4C for 2 weeks at least
  • 23-25 kda
  • Detergents: they all behave similarly in the maltosides (DDM, UDM, DM) and fairly well in CHAPS. they can be exchanged into glucosides too. (The protein looks around 67kD off SEC in DM. Alex has noticed that when he prep the protein in OG, he gets some elution at 67kD, but the major peak in approx 350kD)

Properties:

  • sequence:
MSVAVETFGFFMSALGLLMLGLTLSNSYWRVSTVHGNVITTNTIFENLWYSCATDSLGVSNCWDFPSMLALSGYVQGCRALMITAILLGFLGLFLGMVGLRCTNVGNMDLSKKAKLLAIAGTLHILAGACGMVAISWYAVNITTDFFNPLYAGTKYELGPALYLGWSASLLSILGGICVFSTCCCSSKEEPATRAGLPYKPSTVVIPRATSDESDISFGKYGKNAYVGLVPR

X256 - Dec 6 2013 - Sparse Matrix

Protein in 20mM Tris pH 7.2, 150mM NaCl, 2% glycerol, 0.1% DM: Protein_acquisition_form-cldn15-Dec2013.doc.

Conditions: Sparse matrix + best conditions of CDN3 (3Dx crystals, cf CDN3 twikipage).

Details: X256-CDN15_Sparse.xlsx.

Added:
>
>

X256, results

Image selection: X256-CDN15_ImageSelection.pptx.
 
Added:
>
>		Mostly aggregates (E.coli worse), and small vesicles. 1 striated vesicles (but DOPC lipid, so likely a lipid crystal). Unclear if the protein is even reconstituted in the lipids.
 

  • Set ALLOWTOPICVIEW =

-- NicolasCoudray - 10 Dec 2013

META FILEATTACHMENT attr="" autoattached="1" comment="" date="1386682821" name="Protein_acquisition_form-cldn15-Dec2013.doc" path="Protein_acquisition_form-cldn15-Dec2013.doc" size="60928" user="Main.NicolasCoudray" version="1"
Added:
>
>
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1387304563" name="X256-CDN15_ImageSelection.pptx" path="X256-CDN15_ImageSelection.pptx" size="3874740" user="Main.NicolasCoudray" version="1"
 
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1386682829" name="X256-CDN15_Sparse.xlsx" path="X256-CDN15_Sparse.xlsx" size="587118" user="Main.NicolasCoudray" version="1"

Revision 210 Dec 2013 - Main.NicolasCoudray

 
META TOPICPARENT name="2DCrystallizationExp"

2dx - Claudin-15: 2D crystallization

General 2dx information

  • the protein can be frozen (still leads to 3D crystals)
  • stable at 4C for 2 weeks at least
  • 23-25 kda
Changed:
<
<
  • Detergents: DDM, DM, OG
>
>
  • Detergents: they all behave similarly in the maltosides (DDM, UDM, DM) and fairly well in CHAPS. they can be exchanged into glucosides too. (The protein looks around 67kD off SEC in DM. Alex has noticed that when he prep the protein in OG, he gets some elution at 67kD, but the major peak in approx 350kD)
 

Properties:

  • sequence:
MSVAVETFGFFMSALGLLMLGLTLSNSYWRVSTVHGNVITTNTIFENLWYSCATDSLGVSNCWDFPSMLALSGYVQGCRALMITAILLGFLGLFLGMVGLRCTNVGNMDLSKKAKLLAIAGTLHILAGACGMVAISWYAVNITTDFFNPLYAGTKYELGPALYLGWSASLLSILGGICVFSTCCCSSKEEPATRAGLPYKPSTVVIPRATSDESDISFGKYGKNAYVGLVPR

X256 - Dec 6 2013 - Sparse Matrix

Changed:
<
<		Protein in 20mM Tris pH 7.2, 150mM NaCl?, 2% glycerol, 0.1% DM: Protein_acquisition_form-cldn15-Dec2013.doc.
>
>		Protein in 20mM Tris pH 7.2, 150mM NaCl, 2% glycerol, 0.1% DM: Protein_acquisition_form-cldn15-Dec2013.doc.
 Conditions: Sparse matrix + best conditions of CDN3 (3Dx crystals, cf CDN3 twikipage).

Details: X256-CDN15_Sparse.xlsx.

  • Set ALLOWTOPICVIEW =

-- NicolasCoudray - 10 Dec 2013

META FILEATTACHMENT attr="" autoattached="1" comment="" date="1386682821" name="Protein_acquisition_form-cldn15-Dec2013.doc" path="Protein_acquisition_form-cldn15-Dec2013.doc" size="60928" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1386682829" name="X256-CDN15_Sparse.xlsx" path="X256-CDN15_Sparse.xlsx" size="587118" user="Main.NicolasCoudray" version="1"

Revision 110 Dec 2013 - Main.NicolasCoudray

 
META TOPICPARENT name="2DCrystallizationExp"

2dx - Claudin-15: 2D crystallization

General 2dx information

  • the protein can be frozen (still leads to 3D crystals)
  • stable at 4C for 2 weeks at least
  • 23-25 kda
  • Detergents: DDM, DM, OG

Properties:

  • sequence:
MSVAVETFGFFMSALGLLMLGLTLSNSYWRVSTVHGNVITTNTIFENLWYSCATDSLGVSNCWDFPSMLALSGYVQGCRALMITAILLGFLGLFLGMVGLRCTNVGNMDLSKKAKLLAIAGTLHILAGACGMVAISWYAVNITTDFFNPLYAGTKYELGPALYLGWSASLLSILGGICVFSTCCCSSKEEPATRAGLPYKPSTVVIPRATSDESDISFGKYGKNAYVGLVPR

X256 - Dec 6 2013 - Sparse Matrix

Protein in 20mM Tris pH 7.2, 150mM NaCl?, 2% glycerol, 0.1% DM: Protein_acquisition_form-cldn15-Dec2013.doc.

Conditions: Sparse matrix + best conditions of CDN3 (3Dx crystals, cf CDN3 twikipage).

Details: X256-CDN15_Sparse.xlsx.

  • Set ALLOWTOPICVIEW =

-- NicolasCoudray - 10 Dec 2013

META FILEATTACHMENT attr="" autoattached="1" comment="" date="1386682821" name="Protein_acquisition_form-cldn15-Dec2013.doc" path="Protein_acquisition_form-cldn15-Dec2013.doc" size="60928" user="Main.NicolasCoudray" version="1"
META FILEATTACHMENT attr="" autoattached="1" comment="" date="1386682829" name="X256-CDN15_Sparse.xlsx" path="X256-CDN15_Sparse.xlsx" size="587118" user="Main.NicolasCoudray" version="1"
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