The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/500 - 1/1000. Predicted molecular weight: 93 kDa.Can be blocked with ADAM17 peptide (ab7881). Detects bands of 80-130 kDa bands, which may represent mature protein, precursor, and glycosylated TACE.
Use a concentration of 10 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Use a concentration of 10 µg/ml.
Use at an assay dependent concentration. PubMed: 19553533
ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
Cleaves the membrane-bound precursor of TNF-alpha to its mature soluble form. Responsible for the proteolytical release of soluble JAM3 from endothelial cells surface. Responsible for the proteolytic release of several other cell-surface proteins, including p75 TNF-receptor, interleukin 1 receptor type II, p55 TNF-receptor, transforming growth factor-alpha, L-selectin, growth hormone receptor, MUC1 and the amyloid precursor protein. Also involved in the activation of Notch pathway.
Ubiquitously expressed. Expressed at highest levels in adult heart, placenta, skeletal muscle, pancreas, spleen, thymus, prostate, testes, ovary and small intestine, and in fetal brain, lung, liver and kidney.
Must be membrane anchored to cleave the different substrates. The cytoplasmic domain is not required for the this activity. Only the catalytic domain is essential to shed TNF and p75 TNFR. The conserved cysteine present in the cysteine-switch motif binds the catalytic zinc ion, thus inhibiting the enzyme. The dissociation of the cysteine from the zinc ion upon the activation-peptide release activates the enzyme.
The precursor is cleaved by a furin endopeptidase. Phosphorylated. Stimulation by growth factor or phorbol 12-myristate 13-acetate induces phosphorylation of Ser-819 but decreases phosphorylation of Ser-791.
Predicted band size : 93 kDa Western Blot of HeLa (1, 4), Jurkat (2, 5) and Raji (3, 6) cell lysates labeling ADAM17 with Anti-ADAM17 antibody (ab2051) at 0.5µg/ml in the absence (1-3) or presence of blocking peptide (4-6).
Western blot - Anti-ADAM17 antibody (ab2051)
All lanes : Anti-ADAM17 antibody (ab2051) at 1/500 dilution
Lane 1 : HeLa whole cell lysate with absence of blocking peptide Lane 2 : Jurkat whole cell lysate with absence of blocking peptide Lane 3 : HeLa whole cell lysate with ADAM17 peptide (ab7881) Lane 4 : Jurkat whole cell lysate with ADAM17 peptide (ab7881)
Predicted band size : 93 kDa 80 to 130 kDa bands can be detected, which may represent mature protein, precursor, and glycosylated ADAM17.
IHC image of ADAM17 staining in Human pancreas adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab2051, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
ab2051 at 10µg/ml staining ADAM17 in Hela cells by ICC/IF
Western blot - Anti-ADAM17 antibody (ab2051)This image is courtesy of an anonymous Abreview.
All lanes : Anti-ADAM17 antibody (ab2051) at 1/1000 dilution
Lane 1 : HeLa cell lysate Lane 2 : U937 cell lysate Lane 3 : HT1080 cell lysate Lane 4 : ES cell lysate Lane 5 : L929 cell lysate Lane 6 : CHO cell lysate Lane 7 : 293T cell lysate Lane 8 : ADAM17-overexpressing 293T cell lysate
Lysates/proteins at 50 µg per lane.
Secondary HRP-conjugated Goat anti-Rabbit polyclonal antibody at 1/4000 dilution Developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 93 kDa Observed band size : 93 kDa Additional bands at : 78 kDa (possible isoform).