Mouse Anti-IGF Receptor Monoclonal Antibody from EMD Millipore, used in Immunofluorescence

Primary Antibody: Anti-Insulin-Like Growth Factor (IGF-1) Receptor Monoclonal Antibody, Mouse mAb ( clone alpha-IR3)
Antigen/Epitope: Human IGF-I receptor
Supplier and catalog number: EMD Millipore (previously Calbiochem®) GR11

Sample: 15 µm-thick sections of human colon

Secondary Antibody: TRITC-conjugated AffiniPure Donkey anti-Mouse F(a,b)'2 Fragment (H+L)
Supplier and catalog number: Jackson Immunoresearch 715-026-151

Block: 10% normal donkey serum (NDS) in PBS
Incubation time: 1 hour
Incubation temperature: 25°C

Primary Antibody Incubation Dilution used: 10 ug/ml
Diluent composition: 1% NDS in PBS
Incubation time: 24 hours
Incubation temperature: 4°C

Secondary Antibody Incubation
Dilution used: 1/400
Diluent composition: 1% NDS in PBS
Incubation time: 1 hr
Incubation temperature: 25°C

Detection Method: Confocal microscope (Leica TCS SP2) observation of tissue section mounted onto a slide.

Results Summary:At these concentrations of primary and secondary antibodies, the staining of colon epithelium cells is not very intense, but staining of the chorion is much better.

Additional notes/tips: I pre-incubated sections with 0.15% Triton-X100 in PBS, for 10 min, to permeabilize the cells.