Jun 04, 2025

Public workspaceHuProt (CSF)

DOI
https://dx.doi.org/10.17504/protocols.io.n92ld8pqov5b/v1
  • Jailyn Izu1
  • 1NIMH
Christopher Bartley
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DOI: https://dx.doi.org/10.17504/protocols.io.n92ld8pqov5b/v1
Protocol Citation: Jailyn Izu 2025. HuProt (CSF). protocols.io https://dx.doi.org/10.17504/protocols.io.n92ld8pqov5b/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: May 16, 2024
Last Modified: March 26, 2026
Protocol Integer ID: 99983
Keywords: csf for autoantibody, huprot, autoantibody, screening csf, csf
Funders Acknowledgements:
National Institute of Mental Health
Grant ID: ZIA MH002982
National Institute of Mental Health
Grant ID: R01MH122471
National Institute of Neurological Disorders and Stroke
Grant ID: U01NS120836
Foundation for the National Institutes of Health
Grant ID: Deeda Blair Research Initiative for Disorders of the Brain
Abstract
Screening CSF for autoantibodies by HuProt v4.0
Materials
ReagentBovine Serum Albumin (IgG-Free, Protease-Free)Jackson ImmunoResearch Laboratories, Inc.Catalog #001-000-161
ReagentTWEEN 20Merck MilliporeSigma (Sigma-Aldrich)Catalog #P7949
ReagentAlexa Fluor® 647 AffiniPure™ F(ab)₂ Fragment Goat Anti-Human IgG, Fcγ fragment specificJackson ImmunoResearch Laboratories, Inc.Catalog #109-606-098
ReagentCy™3 AffiniPure™ Goat Anti-Human Serum IgA, α chain specificJackson ImmunoResearch Laboratories, Inc.Catalog #109-165-011

Protocol materials
ReagentTWEEN 20Merck MilliporeSigma (Sigma-Aldrich)Catalog #P7949
ReagentAlexa Fluor® 647 AffiniPure™ F(ab)₂ Fragment Goat Anti-Human IgG, Fcγ fragment specificJackson ImmunoResearch Laboratories, Inc.Catalog #109-606-098
ReagentCy™3 AffiniPure™ Goat Anti-Human Serum IgA, α chain specificJackson ImmunoResearch Laboratories, Inc.Catalog #109-165-011
ReagentBovine Serum Albumin (IgG-Free, Protease-Free)Jackson ImmunoResearch Laboratories, Inc.Catalog #001-000-161
Troubleshooting
Run CSF on HuProt array
Follow the general HuProt microarray protocol.
Protocol
NAME

General HuProt microarray protocol [Nicole Lee]

CREATED BY
Nicole Lee

Protocol-specific parameters:
  • For Steps 1, 19, and 21, the secondary antibodies are:
  1. AlexaFluor 647-cinjugated AffiniPure F(ab')2 Fragment Goat-Antihuman IgG (Catalog number: 109-606-098) ReagentAlexa Fluor® 647 AffiniPure™ F(ab)₂ Fragment Goat Anti-Human IgG, Fcγ fragment specificJackson ImmunoResearch Laboratories, Inc.Catalog #109-606-098
  2. Cy3-conjugated AffiniPure Goat-Antihuman Serum IgA (Catalog number: 109-165-011) ReagentCy™3 AffiniPure™ Goat Anti-Human Serum IgA, α chain specificJackson ImmunoResearch Laboratories, Inc.Catalog #109-165-011

  • For Steps 2 and 4, the sample is 1 mL CSF
  • NOTE for Step 9, Never allow HuProt microarray slides to dry out during this protocol until drying step.