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Our Abpromise guarantee covers the use of ab38611 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 1 - 5 µg/ml.|
|WB||1/1000. Detects a band of approximately 51 kDa (predicted molecular weight: 51 kDa).Can be blocked with Human htrA1 peptide (ab112516).|
|IHC-P||1/10 - 1/50.|
Blocking buffer 5% NFDM/TBST
Diluting buffer 5% NFDM/TBST
ab38611 staining htrA1 in Human liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde, permeabilized with 0.2% Triton X-100 in PBS and blocked with 5% milk for 30 minutes at room temperature; antigen retrieval was by heat mediation in 10mM sodium citrate + 0.05% Tween. Samples were incubated with primary antibody (1/50 in PBS) for 16 hours at 4°C. An Alexa Fluor® 594-conjugated Goat anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling htrA1 with ab38611. A peroxidase-conjugated anti-rabbit IgG was used as the secondary antibody, followed by DAB staining.
Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling htrA1 (green) with ab38611. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG was used as the secondary antibody. Nuclei were stained with DAPI (blue).
ICC/IF image of ab38611 stained HepG2 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab38611, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.