Anti-ADAM17 antibody - Activation site (ab39163)

Overview

  • Product nameAnti-ADAM17 antibody - Activation siteSee all ADAM17 primary antibodies ...
  • Description
    Rabbit polyclonal to ADAM17 - Activation site
  • SpecificityThis antibody recognizes ADAM17, but does not react with other ADAMs.
  • Tested applicationsWB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Pig
  • Immunogen

    Synthetic peptide based on the activation site (cysteine switch and furin cleavage site) of human ADAM17.

    (Peptide available as ab41215.)

  • Epitopeab39163 reacts with an epitope located in the activation site (cysteine switch and furin cleavage site) of ADAM17.

Properties

Applications

Our Abpromise guarantee covers the use of ab39163 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000 - 1/5000. Detects a band of approximately 120 kDa (predicted molecular weight: 93 kDa).Can be blocked with ADAM17 peptide (ab41215). 1/1000 (when using colorimetric substrates such as BCIP/NBT) and 1/5000 (for chemiluminescent substrates). Higher concentrations of antibody may be needed for samples from more distantly related species. Detects a band of approximately 120 kDa in reduced Western blots of conditioned media or cell lysates, which is converted to a 55-60 kDa band.Note: 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 dilution. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent dilution.

Target

  • FunctionCleaves 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.
  • Tissue specificityUbiquitously 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.
  • Sequence similaritiesContains 1 disintegrin domain.
    Contains 1 peptidase M12B domain.
  • DomainMust 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.
  • Post-translational
    modifications
    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.
  • Cellular localizationMembrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • A disintegrin and metalloproteinase domain 17 (tumor necrosis factor, alpha, converting enzyme) antibody
    • A disintegrin and metalloproteinase domain 17 antibody
    • ADA17_HUMAN antibody
    • ADAM 17 antibody
    • ADAM metallopeptidase domain 17 antibody
    • ADAM17 antibody
    • ADAM17 protein antibody
    • CD 156b antibody
    • CD156b antibody
    • CD156b antigen antibody
    • CSVP antibody
    • Disintegrin and metalloproteinase domain-containing protein 17 antibody
    • MGC71942 antibody
    • Snake venom like protease antibody
    • Snake venom-like protease antibody
    • TACE antibody
    • TNF alpha convertase antibody
    • TNF alpha converting enzyme antibody
    • TNF-alpha convertase antibody
    • TNF-alpha-converting enzyme antibody
    • Tumor Necrosis Factor Alpha Converting Enzyme antibody
    see all

Anti-ADAM17 antibody - Activation site images

  • The image shows ab39163 detecting mouse ADAM17 expressed in transfected HeLa cells. The cells were fixed in methanol and incubated with 10 µg/ml ab39163 for 1 hour at 20°C.

  • Anti-ADAM17 antibody - Activation site (ab39163) at 1/4000 dilution + MDA-MB231 lysate

    Secondary
    Donkey anti-Rabbit at 1/10000 dilution

    Predicted band size : 93 kDa

    Image taken from an anonymous Abreview submitted in September 2007

  • Ab39163 staining human normal colon tissue. Staining is localised to cellular membranes.
    Left panel: with primary antibody at 2 ug/ml. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

References for Anti-ADAM17 antibody - Activation site (ab39163)

This product has been referenced in:
  • Chanthaphavong RS  et al. A Role for cGMP in Inducible Nitric-oxide Synthase (iNOS)-induced Tumor Necrosis Factor (TNF) a-converting Enzyme (TACE/ADAM17) Activation, Translocation, and TNF Receptor 1 (TNFR1) Shedding in Hepatocytes. J Biol Chem 287:35887-98 (2012). WB . Read more (PubMed: 22898814) »
  • Takaguri A  et al. A disintegrin and metalloprotease 17 mediates neointimal hyperplasia in vasculature. Hypertension 57:841-5 (2011). IHC-P ; Rat . Read more (PubMed: 21357274) »

See all 7 Publications for this product

Product Wall

Application Western blot
Loading amount 25 µg
Gel Running Conditions Non-reduced Non-Denaturing (Native) (10%)
Sample Mouse Cell lysate - whole cell (hepatocytes)
Specification hepatocytes
Treatment 100 U/ml L-1 treated st various duration
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

Submitted Jun 11 2014

Thank you for your enquiry.

Three human ADAM17 sequences encode proteins with different Cytoplasmic domains, as is the case for other ADAMs proteases. The 824 amino acid sequence encodes a 92,960 Dalton protein, the 807 AA sequence 91,003 Da...

Read More

Thank you very much for your inquiry. The amino end of ADAM-17 is processed by furin-like proteinases during maturation of the protein. The Carboxy end is also cleaved by different proteinases, and the amino end is further cleaved as well. Th...

Read More
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Hela Cells transiently overexpressing mouse ADAM17)
Specification Hela Cells transiently overexpressing mouse ADAM17
Fixative Methanol
Permeabilization No
Username

Abcam user community

Verified customer

Submitted Feb 19 2008

you are correct in assuming that ab39161 recognizes only the prodomain cleaved during ADAM17 maturation. This maturation can be prevented using divalent metal chelators such as EDTA or EGTA. One of the customers who submitted an Abreview for ab39163 no...

Read More

Thank you for your enquiry. Regarding the difference in detected molecular weight, glycosylation and other post-translational modifications increase the apparent molecular weight of ADAM 17 to 120 kDa, and cleavage at the aminoterminal end results ...

Read More
Application Western blot
Sample Human Cell lysate - whole cell (MDA-MB231 Cell Line)
Loading amount 30000 cells
Specification MDA-MB231 Cell Line
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Sep 10 2007

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"