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Our Abpromise guarantee covers the use of ab75732 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||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.
|Flow Cyt||1/10 - 1/50.
ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
|WB||1/1000 - 1/2000. Detects a band of approximately 100 kDa (predicted molecular weight: 100 kDa).|
Immunohistochemical analysis of PFA fixed, paraffin-embedded human brain tissue sections labelling LGR5 with ab75732 at 1/25 dilution.
Blocking buffer 5% NFDM/TBST
Diluting buffer 5% NFDM/TBST
Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling GPCR GPR49 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).
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).
Immunohistochemical analysis of PFA fixed, paraffin-embedded mouse brain tissue sections labelling LGR5 with ab75732 at 1/25 dilution.
Blocking Step: 5% Milk for 1 hour at 20°C.
ab75732 at 0.025 mg/ml staining GPCR GPR49 in human Hela cells by Immunocytochemistry / Immunofluorescence.
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).
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