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Our Abpromise guarantee covers the use of ab28340 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 at an assay dependent concentration. PubMed: 24466060|
|WB||1/1000 - 1/5000. Detects a band of approximately 110-120 kDa (predicted molecular weight: 88 kDa). 1/1000 when using colorimetric substrates such as BCIP/NBT - 1/5000 for chemiluminescent substrates. Bands run high because of post-translational modifications and reduction. EDTA/EGTA treatment of tissues or lysates is required to see latent zymogen. Dilution optimised using Chromogenic detection.|
|IHC-P||Use at an assay dependent concentration.|
ab28340 staining CD26 - Spacer region in rat epididymis tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent paraformaldehyde fixation before heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 and then blocking with 3% hydrogen peroxide in TBS for 10 minutes at 25°C was performed in order to block endogenous peroxidase activity. The primary antibody was used diluted in 1/400 and it was incubated with sample for 16 hours at 4°C in a dilution buffer containing TBS, 0.3% Triton X-100, 0.1 % BSA. A Biotin conjugated goat polyclonal to rabbit IgG was used as secondary antibody at 1/200 dilution.
Immunocytochemistry/Immunofluorscence analysis of IPSC cells labelling CD26 with ab28340 at a dilution of 1/100. Cells were fixed for 30 minutes at 4°C in 4% paraformaldehyde and washed 3 time with DPBS. Cells were blocked for 1 hours with DPBS containing 1% donkey serum and 1% Triton X-100. Cells were incubated for 1 hour at room temperature with ab28340 and ab32572 (anti-beta Catenin, 1/100) in blocking buffer. Cells were washed three times with PBS for 30 minutes each. Cells were incubated for 1 hour at room temperature with appropriate secondary antibodies diluted in the blocking solution. Cells were then washed three times with PBS for 30 minutes each and then imaged using an LSM700 laser scanning confocal microscope.