We are happy to share the protocols and reagents that we have developed for localization of leptospiral proteins.

Bioinformatic Tools

The following are bioinformatic tools that we have found to be useful:

• Identification of Spirochetal Lipoproteins Using SpLip
• The SpLip Algorithm
• PRED-TMBB for Transmembrane Outer Membrane Proteins

Membrane Proteins

Multiple methods should be employed to experimentally verify outer membrane integration and surface exposure.  The following are links to papers describing methods for identification of surface-exposed outer membrane proteins:

• Membrane Integration
• Surface Immunofluorescence
• Surface Biotinylation
• Surface Proteolysis
• Surface Immunoprecipitation
• Surface ELISA

Control Antisera

It is essential to include both positive and negative controls when studying the location of membrane proteins.  We are happy to provide antisera to the following proteins:

Positive Controls

• OmpL54 – a surface-exposed lipoprotein
• OmpL1 – a dominant leptospiral porin

Negative Controls

• FlaA1 – an endoflagellar sheath protein
• LipL31 – an inner membrane lipoprotein
• GroEL – the major cytoplasmic chaparonin

If you would like us to send a small amount of any of these antisera, send an email with your name and mailing address to: David A. Haake

This work is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported License.

Haake Lab Protocols

Coming soon!

• Leptospiral EMJH Culture Medium
• Surface Immunofluorescence Assay
• Surface Proteolysis Assay
• Surface Biotinylation Assay
• Membrane Integration Assay