Institute for Immunity, Transplantation and Infection

The Human Immune Monitoring Center (HIMC)

For more information contact:
Joanna Liliental, Manager
Email » [jlili]

Translational Appliccations Service Ctr (TASC)
Grant Building, Room S107
300 Pasteur Drive
Stanford, CA 94305
tasc.stanford.edu

phone:  650-736-1285

Nanoimmunoassays:  now run through the Translational Applications Service Center (TASC): contact Joanna Liliental (jlili@stanford.edu [jlili]) for details

The NanoPro nanoimmunoassay platform from Protein Simple is an automated microcapillary isoelectric focusing system.  Once focused, proteins are crosslinked to the capillary wall, and subject to antibody probes, much like a Western blot.  Because post-translational events like phosphorylation alter a protein's pI, the system is useful for analyzing signaling events in very small samples.  We have optimized a number of phosphoprotein assays on this system, including Erk1/2, Akt, STAT1, STAT5, and others.

User Instructions

1.  Planning your assay:  Please consult with TASC [jlili] to plan your assay.  There are a number of choices to be made, including lysis conditions, antibody probes, gradient to be used, number of replicates, etc.  See the assay development guide for details.  The NanoPro can process cycles of 12 capillaries at a time, with up to 8 cycles per run.  Replicates (2-4 per sample) are useful in case of technical issues.

2.  Collecting your samples:  See our user guide for details.  Lysates can be frozen at -80C prior to running on the the NanoPro.

3.  Running your samples:  Be sure to plan ahead with TASC [jlili] to coordinate your experiment run.

4.  Data Analysis:  A summary of your data can begenerated using Compass software (Protein Simple).  If needed, futher analysis can be arranged for an additional fee.

Protocols

Stanford Medicine Resources:

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