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

Mouse Anti-IGF Receptor Monoclonal Antibody from EMD Millipore, used in Immunofluorescence
Editor's Note: All antibody reviews appear in protocol format.

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.

AK.Protocol 1.anti-IGF 400.jpg

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

Scientist
Cellular Imaging Department
Biovays
Mouse Anti-IGF Receptor Monoclonal Antibody from EMD Millipore, used in Immunofluorescence
The Good

Clean staining of colon epithelium and chorion.

The Bad

Need to use higher concentrations for more intense staining.

The Bottom Line

Antibody works well in immunofluorescence applications.

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