Lab protocols

Biochemistry protocols

2D Gels (61 KB)
Antibody purification (98 KB)
Silver staining of gels (53 KB)
LPS preparation (94 KB)
LPS removal (68 KB)
OmpC extraction (45 KB)
Pouring protein gels (706 KB)
Running protein gels (811 KB)
Cytoplasmic and membrane protein extraction (38 KB)
Western blot (58 KB)

Cell culture protocols

Generating BM-derived basophils (65 KB)
Generating BM-derived DCs (44 KB)
CFSE labeling of T cells (39 KB)
CFSE proliferation assay for SMARTA T cells (56 KB)
Fusions for mouse hybridomas (105 KB)
Using the hemocytometer (64 KB)
DC isolation from LN (67 KB)
Cryopreservation of primary CD4+ T cells (60 KB)

Cell isolation protcols

Positive cell enrichment by MACS (63 KB)
Colonic lamina propria preparation (104 KB)
Small intestinal lamina propria preparation (76 KB)

ELISA and CBA protocols

Albumin ELISA (88 KB)
Antigen-specific ELISA_bacterial cytoplasmic proteins (66 KB)
Dirty-plate ELISA (47 KB)
Ig ELISA using mouse serum, intestinal washes and fragment cultures (171 KB)
IL-2 specific ELISA (185 KB)
Limulus assay (90 KB)
Total and anti-Remicade ELISA (55 KB)
BD cytometric bead array (204 KB)
IgE ELISA for mouse serum (298 KB)

Flow cytometry

Microbiology protocols

16S genomic DNA isolation (55 KB)
16S PCR protocol (59 KB)
Bacterial plating (109 KB)
Making electro-competent cells (16 KB)
Culturing anaerobic bacteria from frozen stocks (197 KB)
Electroporation (78 KB)
Gavagingbacteria (75 KB)

Encyclopedia of microbiology

Encyclopedia of Microbiology (3rd Edition), Elsevier. Pages 237-246.
Gnotobiotic and Axenic Animals.
Macpherson AJ, Geuking M.B., Kirundi J, Collins S, and McCoy KD. 2009.
Link to article - PDF (password)

Protocol for LB agar plates (42 KB)
Summary of autoclave programs (57 KB)
Protocol for bacterial sampling from mouse intestine (47 KB)
Protocol for importation of food after setting up an isolator (50 KB)
Protocol for importation of material after setting up an isolator (53 KB)
Protocol for importation of sawdust into an isolator (49 KB)
Protocol for importation of water supplies after setting up an isolator (57 KB)
Summary of media recipes (37 KB)
Protocol for setup of germ-free isolator (49 KB)
Protocols of bacterial culture methods for sterility testing of samples from the gnotobiotic unit (90 KB)
Sytox-green staining of intestinal content (42 KB)
Germ-free vasectomy of mice (68 KB)