Product datasheet
Anti-ADAM17 antibody [EPR3600(N)(B)] (Alexa Fluor®
488) ab196457
2 Images
Overview
Product name
Anti-ADAM17 antibody [EPR3600(N)(B)] (Alexa Fluor® 488)
Description
Rabbit monoclonal [EPR3600(N)(B)] to ADAM17 (Alexa Fluor® 488)
Conjugation
Alexa Fluor® 488. Ex: 495nm, Em: 519nm
Tested applications
ICC/IF, Flow Cyt
Species reactivity
Reacts with: Mouse, Rat, Human
Immunogen
Synthetic peptide within Human ADAM17 aa 750 to the C-terminus (Cysteine residue). The
exact sequence is proprietary.
Database link: P78536
Positive control
ICC/IF: HeLa cells. Flow Cyt: HeLa cells.
General notes
This product is a recombinant rabbit monoclonal antibody.
Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the
following U.S. Patents, No. 5,675,063 and/or 7,429,487.
.
Alternative versions available:
Anti-ADAM17 antibody [EPR3600(N)(B)] (ab173579)
Anti-ADAM17 antibody (Alexa Fluor® 647) [EPR3600(N)(B)] (ab196581)
Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific
Company. The Alexa Fluor® dye included in this product is provided under an intellectual
property license from Life Technologies Corporation. As this product contains the Alexa Fluor®
dye, the purchase of this product conveys to the buyer the non-transferable right to use the
purchased product and components of the product only in research conducted by the buyer
(whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor®
dye the sale of this product is expressly conditioned on the buyer not using the product or its
components, or any materials made using the product or its components, in any activity to
generate revenue, which may include, but is not limited to use of the product or its components:
in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for
therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they
are sold for use in research. For information on purchasing a license to use products containing
Alexa Fluor® dyes for purposes other than research, contact Life Technologies Corporation,
5791 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@lifetech.com.
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Properties
Form
Liquid
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long
term. Stable for 12 months at -20°C. Store In the Dark.
Storage buffer
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: PBS, 30% Glycerol, 1% BSA
Purity
Immunogen affinity purified
Clonality
Monoclonal
Clone number
EPR3600(N)(B)
Isotype
IgG
Applications
Our Abpromise guarantee covers the use of ab196457 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application
Abreviews
ICC/IF
Notes
1/50.
This product gave a positive signal in both 100% methanol (5 min) and 4%
formaldehyde (10 min) fixed HeLa cells.
Flow Cyt
1/50.
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 activationpeptide 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.
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Anti-ADAM17 antibody [EPR3600(N)(B)] (Alexa Fluor® 488) images
Overlay histogram showing HeLa cells
stained with ab196457 (red line). The cells
were fixed with 80% methanol (5 min) and
then permeabilized with 0.1% PBS-Tween for
20 min. The cells were then incubated in 1x
PBS / 10% normal goat serum / 0.3M glycine
to block non-specific protein-protein
interactions followed by the antibody
(ab196457, 1/50 dilution) for 30 min at 22°C.
Isotype control antibody (black line) was
rabbit monoclonal IgG [EPR25A] Alexa
Flow Cytometry - Anti-ADAM17 antibody
Fluor® 488 (ab199091) used at the same
[EPR3600(N)(B)] (Alexa Fluor® 488) (ab196457)
concentration and conditions as the primary
antibody. Unlabelled sample (blue line) was
also used as a control. Acquisition of >5,000
events were collected using a 20mW Argon
ion laser (488nm) and 525/30 bandpass filter.
ab196457 staining ADAM17 in HeLa cells.
The cells were fixed with 4% formaldehyde
(10 min), permeabilized in 0.1% Triton X-100
for 5 minutes and then blocked in 1%
BSA/10% normal goat serum/0.3M glycine in
0.1% PBS-Tween for 1h. The cells were then
incubated with ab196457 at 1/50
dilution (shown in green) and ab195889,
Mouse monoclonal [DM1A] to alpha Tubulin
(Alexa Fluor® 594, shown in red) at 2µg/ml
overnight at +4°C. Nuclear DNA was labelled
Immunocytochemistry/ Immunofluorescence -
in blue with DAPI.
Anti-ADAM17 antibody [EPR3600(N)(B)] (Alexa
Fluor® 488) (ab196457)
This product gave a positive signal in 100%
methanol (5 min) fixed SKNSH cells under the
same testing conditions.
Image was taken with a confocal microscope
(Leica-Microsystems, TCS SP8).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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Replacement or refund for products not performing as stated on the datasheet
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