Overview

  • Product name
  • Description
    Rabbit polyclonal to ADAM17
  • Host species
    Rabbit
  • Specificity
    This antibody recognizes ADAM17, but does not react with other ADAMs. ab39163 reacts with an epitope located in the activation site (cysteine switch and furin cleavage site) of ADAM17.
  • Tested applications
    Suitable for: WB, 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.

    Read Abcam's proprietary immunogen policy (Peptide available as ab41215.)

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). 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 concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent concentration.

Target

  • Function
    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.
  • Tissue specificity
    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.
  • Sequence similarities
    Contains 1 disintegrin domain.
    Contains 1 peptidase M12B domain.
  • Domain
    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.
  • 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 localization
    Membrane.
  • 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
    • NISBD antibody
    • NISBD1 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

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 (ab39163) at 1/4000 dilution + MDA-MB231 lysate

    Secondary
    Donkey anti-Rabbit at 1/10000 dilution

    Predicted band size: 93 kDa



    ab39163 detected ADAM17 protein in lysates of the highly invasive human breast cancer cell line MDA-MB231. ab39163 was incubated at a dilution of 1/4000 for 16 hours at 4°C in PBS + 5% Milk. For further details please refer to the Abreview.

  • 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

This product has been referenced in:
  • Oikonomidi I  et al. iTAP, a novel iRhom interactor, controls TNF secretion by policing the stability of iRhom/TACE. Elife 7:N/A (2018). Read more (PubMed: 29897333) »
  • Zocchi MR  et al. ADAM10 new selective inhibitors reduce NKG2D ligand release sensitizing Hodgkin lymphoma cells to NKG2D-mediated killing. Oncoimmunology 5:e1123367 (2016). Read more (PubMed: 27467923) »
See all 14 Publications for this product

Customer reviews and Q&As

1-10 of 11 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Skin - Melanoma)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: EDTA pH = 8.0
Permeabilization
No
Specification
Skin - Melanoma
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
Paraformaldehyde

Dr. Christian Ostalecki

Verified customer

Submitted Aug 07 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Intestine and skin)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris-EDTA
Permeabilization
No
Specification
Intestine and skin
Fixative
Formaldehyde

Dr. Pilar Alzuguren

Verified customer

Submitted Aug 01 2016

Application
Immunoprecipitation
Sample
Mouse Cell lysate - whole cell (mouse hepatocytes)
Total protein in input
300 µg
Treatment
100 ng/ml LPS
Immuno-precipitation step
Protein A/G
Specification
mouse hepatocytes

Abcam user community

Verified customer

Submitted Nov 27 2015

Application
Western blot
Sample
Human Cell lysate - whole cell (human hepatocytes)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Treatment
100 ng/ml LPS
Specification
human hepatocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Sep 09 2015

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

Abcam user community

Verified customer

Submitted Jun 11 2014

Question
Answer

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 Dalton, and the 694 AA sequence 78,543 Dalton predicted MW. Glycosylation and other posttranslational modifications increase the apparent molecular weight on PAGE gels. Mouse and rat ADAM17 sequences are both 827 AA, predicting 93,073 and 93,017 Dalton proteins, respectively. This product recognizes a 120 kD band in reduced Western blots of conditioned media or cell lysates, which is converted to a 55-60 kD band.

Read More

Answer

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. The ab39163 ADAM-17 epitope is after the furin cleavage site, within the range of the catalytic domain. Most of the glycosylation occurs in the propeptide domain, so a larger than expected size might be ADAM-17 with intact carboxyterminus. This has not been experimentally tested. I hope this information is helpful. Please do not hesitate to contact me again with any further questions.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Hela Cells transiently overexpressing mouse ADAM17)
Specification
Hela Cells transiently overexpressing mouse ADAM17
Fixative
Methanol
Permeabilization
No

Abcam user community

Verified customer

Submitted Feb 19 2008

Answer

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 noted that "Seems to be selective for Precursor forms of ADAM17". The presence of the 55kDa band as well would indicate that both the cleaved and uncleaved form of this protein are recognized.

Read More

Answer

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 in lower molecular weight forms, therefore producing the 55-60 kDa range. If there is anything else I can help you with, please do not hesitate to contact me.

Read More

1-10 of 11 Abreviews or Q&A

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

Sign up