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Synthetic peptide derived from within residues 950 to the C-terminus of Human Insulin degrading enzyme/ IDE.
(Peptide available as ab32215.)
Our Abpromise guarantee covers the use of ab32216 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 118 kDa (predicted molecular weight: 118 kDa).Can be blocked with Human Insulin degrading enzyme / IDE peptide (ab32215).|
|IHC-P||1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Insulin degrading enzyme / IDE knockout HAP1 cell lysate (20 µg)
Lane 3: K562 cell lysate (20 µg)
Lane 4: HepG2 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab32216 observed at 120 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab32216 was shown to specifically react with Insulin degrading enzyme / IDE when Insulin degrading enzyme / IDE knockout samples were used. Wild-type and Insulin degrading enzyme / IDE knockout samples were subjected to SDS-PAGE. ab32216 and ab8245 (loading control to GAPDH) were diluted at 1 μg/ml and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 h at room temperature before imaging.
Image courtesy of Human Protein Atlas
ab32216 staining human Gall bladder. The paraffin embedded human skin tissue was incubated with ab32216 (1/50 dilution) for 30 mins at room temperature. The staining is predominantly cytoplasmic although some nuclear staining can be seen. Antigen retrieval was performed by heat induction in citrate buffer pH 6. Ab32216 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines.
Further results for
ab32216 staining cultured rat OCP by ICC/IF. The cultured cells were fixed with paraformaldehyde and blocked with 10% donkey serum for 30 minutes at 24°C. The cultured cellswere then stained with ab322161 at 1/500 in 0.3% TritonX with 0.1% PBS and 10% donkey serum for 4h at 24°C. An Alexa Fluro 488 donkey anti-rabbit polyclonal antibody at 1/1000 was used as the secondary antibody. Nuclei were stained with 1.43µM Hoechst and can be observed in blue.