LGR5 is a multi-pass membrane protein, due to hydrophobic nature these proteins tend to aggregate when boiled at 100°C in western blotting therefore, we recommend slow heating of lysates for 15-20 minutes at 60-70°C rather than boiling.
The immunogen used for this product shares 62% homology with LGR6. Cross-reactivity with this protein has not been confirmed experimentally.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/10 - 1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Please note for better results the antigen retrieval method may need optimizations. We recommend trying a range of incubation time e.g. 1, 2, 3, 4, 5 minutes using pressure cooker or 3, 5, 10, 15 minutes in microwave after buffer start boiling. For more details check IHC-P – detailed guide in protocol section.
1/10 - 1/50.
ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
1/1000 - 1/2000. Detects a band of approximately 100 kDa (predicted molecular weight: 100 kDa).Can be blocked with Human LGR5 peptide (ab185636).
Orphan receptor. Stem cell marker of the intestinal epithelium and the hair follicule. Target gene of Wnt signaling.
Expressed in skeletal muscle, placenta, spinal cord, and various region of brain. Expressed at the base of crypts in colonic and small mucosa stem cells. In premalignant cancer expression is not restricted to the cript base. Overexpressed in cancers of the ovary, colon and liver.
Belongs to the G-protein coupled receptor 1 family. Contains 16 LRR (leucine-rich) repeats. Contains 1 LRRNT domain.
Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling LGR5 with ab75732. Cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min) and then incubated with the primary antibody (1/100, 2 h at room temperature). An Alexa Fluor® 488 conjugated donkey anti-rabbit antibody (green) was used as the secondary antibody (1/1000, 1h). Nuclei were counterstained with Hoechst 33342 (blue) (10 μg/ml, 5 min).
Flow Cytometry - Anti-LGR5 antibody (ab75732)
Overlay histogram showing 2% paraformaldehyde fixed SH-SY5Y cells stained with ab75732 (green line). Cells were incubated with the antibody at 1/25 dilution at 37oC for 60 minutes. The secondary antibody used was DyLight® 488 Goat-Anti-Rabbit IgG at 1/400 dilution. A non-specific IgG antibody was used for an isotype control (blue line).
ab75732 at 0.025 mg/ml staining LGR5 in human Hela cells by Immunocytochemistry / Immunofluorescence.
Flow Cytometry - Anti-LGR5 antibody (ab75732)This image is courtesy of an anonymous abreview.
Overlay histogram showing mouse intestinal cells stained with ab75732 (pink line). Cells were incubated with the antibody at 1/50 in DMEM at 20°C for 30 minutes.The secondary antibody used was DyLight® 649 goat anti-rabbit IgG at 1/100 dilution. A non-specific IgG antibody was used for an isotype control (black line).