Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-ADAM10 antibody [EPR5622] (ab124695)

Overview

  • Product name

    Anti-ADAM10 antibody [EPR5622]
    See all ADAM10 primary antibodies
  • Description

    Rabbit monoclonal [EPR5622] to ADAM10
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human ADAM10 (C terminal). The exact sequence is proprietary.

  • Positive control

    • Jurkat, RAW 264.7, U2OS, LnCaP and MCF-7 cell lysates.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab124695 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 - 1/10000. Predicted molecular weight: 84 kDa.
IP 1/10 - 1/100.

Target

  • Function

    Cleaves the membrane-bound precursor of TNF-alpha at '76-Ala-
    -Val-77' 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 heparin-binding epidermal growth-like factor, ephrin-A2 and for constitutive and regulated alpha-secretase cleavage of amyloid precursor protein (APP). Contributes to the normal cleavage of the cellular prion protein. Involved in the cleavage of the adhesion molecule L1 at the cell surface and in released membrane vesicles, suggesting a vesicle-based protease activity. Controls also the proteolytic processing of Notch and mediates lateral inhibition during neurogenesis.
  • Tissue specificity

    Expressed in spleen, lymph node, thymus, peripheral blood leukocyte, bone marrow, cartilage, chondrocytes and fetal liver.
  • Sequence similarities

    Contains 1 disintegrin domain.
    Contains 1 peptidase M12B domain.
  • Domain

    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.
  • Cellular localization

    Cell membrane. Endomembrane system. Is localized in the plasma membrane but is predominantly expressed in the Golgi apparatus and in released membrane vesicles derived likely from the Golgi.
  • Information by UniProt
  • Database links

  • Alternative names

    • A disintegrin and metalloprotease domain 10 antibody
    • A disintegrin and metalloproteinase domain 10 antibody
    • AD 10 antibody
    • AD10 antibody
    • AD18 antibody
    • ADA10_HUMAN antibody
    • ADAM 10 antibody
    • ADAM metallopeptidase domain 10 antibody
    • ADAM10 antibody
    • CD 156c antibody
    • CD156c antibody
    • CD156c antigen antibody
    • CDw156 antibody
    • disintegrin and metalloproteinase domain containing protein 10 antibody
    • Disintegrin and metalloproteinase domain-containing protein 10 antibody
    • HsT 18717 antibody
    • HsT18717 antibody
    • Kuz antibody
    • Kuzbanian antibody
    • Kuzbanian protein homolog antibody
    • Kuzbanian, Drosophila, homolog of antibody
    • MADM antibody
    • Mammalian disintegrin metalloprotease antibody
    • Mammalian disintegrin-metalloprotease antibody
    • RAK antibody
    see all

Images

  • Lane 1: Wild type HAP1 whole cell lysate (40 µg)
    Lane 2: Empty lane
    Lane 3: ADAM10 knockout HAP1 whole cell lysate (40 µg)
    Lane 4: U20S whole cell lysate (40 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab124695 observed at 90 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab124695 was shown to recognize ADAM10 when ADAM10 knockout samples were used, along with additional cross-reactive bands. Wild-type and ADAM10 knockout samples were subjected to SDS-PAGE.  Ab124695 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • ab124695 (purified) at 1:30 dilution (2ug) immunoprecipitating ADAM-10 in LNCaP (Human prostate carcinoma epithelial cell) whole cell lysate 10ug.
    Lane 1 (input): LNCaP (Human prostate carcinoma epithelial cell) whole cell lysate 10ug
    Lane 2 (+): ab124695 & LNCaP (Human prostate carcinoma epithelial cell) whole cell lysate 10ug
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab170952 in HeLa LNCaP (Human prostate carcinoma epithelial cell) whole cell lysateFor western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.

  • All lanes : Anti-ADAM10 antibody [EPR5622] (ab124695) at 0.05 µg/ml (Purified)

    Lane 1 : RAW264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
    Lane 2 : Rat brain lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 84 kDa



    Blocking and diluting buffer: 5% NFDM/TBST

  • All lanes : Anti-ADAM10 antibody [EPR5622] (ab124695) at 0.09 µg/ml (Purified)

    Lane 1 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate
    Lane 2 : MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysate
    Lane 3 : LNCaP (Human prostate carcinoma epithelial cell) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 84 kDa



    Blocking and diluting buffer: 5% NFDM/TBST

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD
  • All lanes : Anti-ADAM10 antibody [EPR5622] (ab124695) at 1/1000 dilution

    Lane 1 : Jurkat cell lysate
    Lane 2 : RAW 264.7 cell lysate
    Lane 3 : LnCaP cell lysate
    Lane 4 : MCF-7 cell lysate

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 84 kDa



    84 kDa band is pro-ADAM10, smaller band at around 65 kDa is mature ADAM10 protein.

References

This product has been referenced in:

  • Li H  et al. Long non-coding RNA CDKN2B-AS1 reduces inflammatory response and promotes cholesterol efflux in atherosclerosis by inhibiting ADAM10 expression. Aging (Albany NY) 11:1695-1715 (2019). Read more (PubMed: 30926762) »
  • Brummer T  et al. NrCAM is a marker for substrate-selective activation of ADAM10 in Alzheimer's disease. EMBO Mol Med 11:N/A (2019). Read more (PubMed: 30833305) »
See all 15 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa)
Gel Running Conditions
Reduced Denaturing (10% gel)
Loading amount
15 µg
Specification
HeLa
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Aug 14 2019

Application
Immunohistochemistry free floating
Sample
Mouse Tissue sections (PFA fixed Brain)
Specification
PFA fixed Brain

Abcam user community

Verified customer

Submitted Feb 14 2019

Application
Western blot
Sample
Rat Cell lysate - whole cell (primary astrocytes)
Loading amount
1 µg
Specification
primary astrocytes
Treatment
LPS 0.1ug/ml, 6h
Gel Running Conditions
Reduced Denaturing (10% gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Apr 16 2013

Answer

The Anti-ADAM10 antibody [EPR5622] (ab124695) has been raised against a synthetic peptide taken from within the region 700 to the C-terminus of the human ADAM10 protein (Uniprot reference O14672).

Read More

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