ITN Protocols

The Immune Tolerance Network (ITN) provides the standard operating procedures and protocols, developed by ITN personnel and collaborators, free to use by the research community. Please note that no technical support is available. All works are licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 2.5 License. 

Detailed Protocols

Following are the standard operating procedures and protocols developed and used by the ITN.

Cell Assay

001
Whole Blood Basophil Activation Assay
v.01.2
ELISPOT 001
Performing Alloreactive ELISPOT Assays
v.001
ITN 2800
Lymphocyte Proliferation Assay Using 3H-Thymidine Incorporation
v.001
P1 RD 011
Detection of IFNg and IL-10 Secreting Cells by ELISPOT
v.001

Cell/Tissue Isolation and Prep

000
Recovery of Frozen Mononuclear Cells
v.002
000
Separation and Freezing Technique for Spleen Mononuclear Cells
v.01.8
000
Thawing Frozen Mononuclear Cells
v.001
KC SOP 001
Collection and Preservation of Kidney or Liver Transplant Biopsy
v.002
LN1
Separation and Freezing Technique for Lymph Node Mononuclear Cells
v.001
PBMC CPT
Isolation and Cryopreservation of PBMC – CPT without Plasma Collection
v.009
PBMC Ficoll
Separation of Mononuclear Cells from Whole Blood Using Ficoll Gradients
v.08.1

Flow Cytometry

ITI 00046
Whole Blood Washing to Remove Components that Interfere with Ab Binding
v.001

Flow Cytometry Panels

FCP01
APC 1 Flow Panel
v.001
FCP02
APC 2 Flow Panel
v.001
FCP03
B Cell Flow Panel
v.001
FCP04
Modified X-Trial T Cell Flow Panel
v.001
FCP05
T Cell Exhaustion Flow Panel
v.001
FCP06
Tfh/lin 1 Flow Panel
v.001
FCP07
Tfh/lin 2 Flow Panel
v.001
FCP08
Treg Flow Panel
v.001
FCP09
X-Trial T Cell Flow Panel
v.001

RNA/DNA

000
Synthesizing cDNA Using ThermoScript RT-PCR Kit in 96 Plate Format
v.001
000
Synthesizing cDNA Using ThermoScript RT-PCR Kit
v.001
RNA 004
Assessing the quality of RNA using the Agilent 2100 Bioanalyzer
v.003