ADAM10 Antibody

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ProSci
Product Code: PSI-2051
Product Group: Primary Antibodies
Supplier: ProSci
CodeSizePrice
PSI-2051-0.02mg0.02mg£150.00
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PSI-2051-0.1mg0.1mg£449.00
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Overview

Host Type: Rabbit
Antibody Isotype: IgG
Antibody Clonality: Polyclonal
Regulatory Status: RUO
Applications:
  • Enzyme-Linked Immunosorbent Assay (ELISA)
  • Immunofluorescence (IF)
  • Immunohistochemistry (IHC)
  • Western Blot (WB)

Images

1 / 15
<strong>Figure 1 ADAM10 KO Validation in MEF Cells </strong><br> Loading: 10 μg of lysate Antibodies: ADAM10 2051, 1 μg/mL and beta-actin 3779-1301, 1μg/mL, 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution. 2051 detected both precursor ADAM10 (94KD) and mature ADAM10 (68kD).
2 / 15
<strong>Figure 2 ADAM10 KO Validation in 293 Cells </strong><br> Loading: 15 μg of lysate Antibodies: ADAM10 2051, 2 μg/mL and beta-actin 3779-1301, 1μg/mL, 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution. 2051 detected both precursor ADAM10 (94KD) and mature ADAM10 (68kD).
3 / 15
<strong>Figure 3 Independent Antibody Validation (IAV) via Protein Expression Profile in Human and Mouse Cell Lines</strong><br> Loading: 15 μg of lysates per lane. Antibodies: ADAM10 2051, 0.5 μg/mL, ADAM10 24-024, 1 μg/mL, and ?-actin 3779, 1 μg/mL, 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
4 / 15
<strong>Figure 4 WB Validation in Human Cell Lines </strong><br> Loading: 15 μg of lysate Antibodies: ADAM10 2051, 1 μg/mL, 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution. 2051 detected both precursor ADAM10 (94KD) and mature ADAM10 (68kD).
5 / 15
<strong>Figure 5 Western Blot Validation in Mouse Tissues </strong><br> Loading: 15 μg of lysates per lane. Antibodies: ADAM10 2051, 1 μg/mL, 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. 2051 detected both precursor ADAM10 (94KD) and mature ADAM10 (68kD).
6 / 15
<strong>Figure 6 Western Blot Validation in Rat Tissues</strong><br> Loading: 15 μg of lysates per lane. Antibodies: ADAM10 2051, 1 μg/mL, 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. 2051 detected both precursor ADAM10 (94KD) and mature ADAM10 (68kD).
7 / 15
<strong>Figure 7 ImmunoFluor®scence Validation of ADAM10 in MOLT4 Cells</strong><br> ImmunoFluor®scent analysis of 4% paraformaldehyde-fixed MOLT4 cells labeling ADAM10 with 2051 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
8 / 15
<strong>Figure 8 ImmunoFluor®scence Validation of ADAM10 in K562 Cells</strong><br> ImmunoFluor®scent analysis of 4% paraformaldehyde-fixed K562 cells labeling ADAM10 with 2051 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
9 / 15
<strong>Figure 9 ImmunoFluor®scence Validation of ADAM10 in Mouse Testis</strong><br> ImmunoFluor®scent analysis of 4% paraformaldehyde-fixed mouse testis labeling ADAM10 with 2051 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
10 / 15
<strong>Figure 10 ImmunoFluor®scence Validation of ADAM10 in Rat Testis </strong><br> ImmunoFluor®scent analysis of 4% paraformaldehyde-fixed rat testis labeling ADAM10 with 2051 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
11 / 15
<strong>Figure 11 WB Validation of ADAM10 in Human washed platelets (Al-Tamimi et al., 2012) </strong><br> Human washed platelets (5x 108/mL) were lysed and ADAM10 expression was detected by anti-ADAM10 cytoplasmic tail antibody (ADAM10 CT). Only mature ADAM10 (~72kD), when got cleaved upon activation, was detectable in resting platelet lysates. Vertical lines indicate a repositioned lane.
12 / 15
<strong>Figure 12 Regulated Expression Validation of ADAM10 in Rat cortical cultures (Hurtado et al., 2002) </strong><br> Western Blot analysis was carried out to monitor protein expression of ADAM10 with anti-ADAM10 antibodies. ADAM10 expression was decreased in rat cortical cultures when exposed to oxygen-glucose deprivation (OGD ) or glutamate.
13 / 15
<strong>Figure 13 ImmunoFluor®scence Validation of ADAM10 in primary cultures of human cerebral vascular smooth muscle cells (HC-VSMCs) (Coma et al., 2008)</strong><br> Detection of ADAM10 expression by anti-ADAM10 antibodies in HC-VSMC cells under control condition or in the presence of 10μM H2O2 (oxidative stress condition) for 6 h. ADAM10 expression was not affected when exposed to oxidative stress.
14 / 15
<strong>Figure 14 Regulated Expression Validation of ADAM10 in Human neuroblastoma (SH-SY5Y) cells (Zimmermann et al., 2004) </strong><br> Protein expression of ADAM10 detected by anti-ADAM10 CT antibodies in control or donepezil treated SH-SY5Y cells. When treated with donepezil, the expression of mature form of ADAM10 (68kD) was up-regulated in membrane compartment as compared to the down-regulation in intracellular fractions, and was not affected in whole cell homogenate.
15 / 15
<strong>Figure 15 KD Validation of ADAM10 in Human embryonic kidney 293 cells overexpressing the human APP 695 isoform (HEK/APP) (Gatta et al., 2009) </strong><br> Western blot analysis of ADAM10 silencing using anti-ADAM10 antibodies in HEK/APP cells. Silencing with ADAM10 siRNA (Ad) significantly decreased ADAM10 expression, and so did with Ferrochelatase siRNA (F) and N-methylprotoporphyrin IX siRNA (N), 67% and 50% reduction respectively
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<strong>Figure 1 ADAM10 KO Validation in MEF Cells </strong><br> Loading: 10 μg of lysate Antibodies: ADAM10 2051, 1 μg/mL and beta-actin 3779-1301, 1μg/mL, 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution. 2051 detected both precursor ADAM10 (94KD) and mature ADAM10 (68kD).
<strong>Figure 2 ADAM10 KO Validation in 293 Cells </strong><br> Loading: 15 μg of lysate Antibodies: ADAM10 2051, 2 μg/mL and beta-actin 3779-1301, 1μg/mL, 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution. 2051 detected both precursor ADAM10 (94KD) and mature ADAM10 (68kD).
<strong>Figure 3 Independent Antibody Validation (IAV) via Protein Expression Profile in Human and Mouse Cell Lines</strong><br> Loading: 15 μg of lysates per lane. Antibodies: ADAM10 2051, 0.5 μg/mL, ADAM10 24-024, 1 μg/mL, and ?-actin 3779, 1 μg/mL, 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 4 WB Validation in Human Cell Lines </strong><br> Loading: 15 μg of lysate Antibodies: ADAM10 2051, 1 μg/mL, 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution. 2051 detected both precursor ADAM10 (94KD) and mature ADAM10 (68kD).
<strong>Figure 5 Western Blot Validation in Mouse Tissues </strong><br> Loading: 15 μg of lysates per lane. Antibodies: ADAM10 2051, 1 μg/mL, 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. 2051 detected both precursor ADAM10 (94KD) and mature ADAM10 (68kD).
<strong>Figure 6 Western Blot Validation in Rat Tissues</strong><br> Loading: 15 μg of lysates per lane. Antibodies: ADAM10 2051, 1 μg/mL, 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. 2051 detected both precursor ADAM10 (94KD) and mature ADAM10 (68kD).
<strong>Figure 7 ImmunoFluor®scence Validation of ADAM10 in MOLT4 Cells</strong><br> ImmunoFluor®scent analysis of 4% paraformaldehyde-fixed MOLT4 cells labeling ADAM10 with 2051 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
<strong>Figure 8 ImmunoFluor®scence Validation of ADAM10 in K562 Cells</strong><br> ImmunoFluor®scent analysis of 4% paraformaldehyde-fixed K562 cells labeling ADAM10 with 2051 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
<strong>Figure 9 ImmunoFluor®scence Validation of ADAM10 in Mouse Testis</strong><br> ImmunoFluor®scent analysis of 4% paraformaldehyde-fixed mouse testis labeling ADAM10 with 2051 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
<strong>Figure 10 ImmunoFluor®scence Validation of ADAM10 in Rat Testis </strong><br> ImmunoFluor®scent analysis of 4% paraformaldehyde-fixed rat testis labeling ADAM10 with 2051 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
<strong>Figure 11 WB Validation of ADAM10 in Human washed platelets (Al-Tamimi et al., 2012) </strong><br> Human washed platelets (5x 108/mL) were lysed and ADAM10 expression was detected by anti-ADAM10 cytoplasmic tail antibody (ADAM10 CT). Only mature ADAM10 (~72kD), when got cleaved upon activation, was detectable in resting platelet lysates. Vertical lines indicate a repositioned lane.
<strong>Figure 12 Regulated Expression Validation of ADAM10 in Rat cortical cultures (Hurtado et al., 2002) </strong><br> Western Blot analysis was carried out to monitor protein expression of ADAM10 with anti-ADAM10 antibodies. ADAM10 expression was decreased in rat cortical cultures when exposed to oxygen-glucose deprivation (OGD ) or glutamate.
<strong>Figure 13 ImmunoFluor®scence Validation of ADAM10 in primary cultures of human cerebral vascular smooth muscle cells (HC-VSMCs) (Coma et al., 2008)</strong><br> Detection of ADAM10 expression by anti-ADAM10 antibodies in HC-VSMC cells under control condition or in the presence of 10μM H2O2 (oxidative stress condition) for 6 h. ADAM10 expression was not affected when exposed to oxidative stress.
<strong>Figure 14 Regulated Expression Validation of ADAM10 in Human neuroblastoma (SH-SY5Y) cells (Zimmermann et al., 2004) </strong><br> Protein expression of ADAM10 detected by anti-ADAM10 CT antibodies in control or donepezil treated SH-SY5Y cells. When treated with donepezil, the expression of mature form of ADAM10 (68kD) was up-regulated in membrane compartment as compared to the down-regulation in intracellular fractions, and was not affected in whole cell homogenate.
<strong>Figure 15 KD Validation of ADAM10 in Human embryonic kidney 293 cells overexpressing the human APP 695 isoform (HEK/APP) (Gatta et al., 2009) </strong><br> Western blot analysis of ADAM10 silencing using anti-ADAM10 antibodies in HEK/APP cells. Silencing with ADAM10 siRNA (Ad) significantly decreased ADAM10 expression, and so did with Ferrochelatase siRNA (F) and N-methylprotoporphyrin IX siRNA (N), 67% and 50% reduction respectively

Documents

Further Information

Additional Names:
ADAM10 Antibody: RAK, kuz, AD10, AD18, MADM, CD156c, HsT18717, KUZ, Disintegrin and metalloproteinase domain-containing protein 10, CDw156, ADAM 10
Application Note:
ADAM10 can be used for detection of ADAM10 by Western blot at 1 - 2 μg/mL. This polyclonal antibody can also detect ADAM10 by immunohistochemistry at 2 μg/mL. For immunofluorescence start at 10 μg/mL.

Antibody validated: Western Blot in human samples; Immunocytochemistry in human samples and Immunofluorescence in human samples. All other applications and species not yet tested.
Background:
ADAM10 Antibody: Proinflammatory cytokine tumor necrosis factor-alpha (TNF-α) contributes to a variety of inflammatory responses and programmed cell death. Notch receptor and its ligand participate in cell fate decisions during vertebrate development and are associated with several human disorders, including a T-cell lymphoma. TNF-α, notch and its ligand delta are all membrane-bond molecules, which are cleaved by proteases to release mature proteins or functional receptor. ADAM10, a metalloprotease-disintegrin in the family of mammalian ADAM (for a disintegrin and metalloprotease), was recently identified to cleave TNF-α, notch and its ligand delta. The genes encoding human, mouse, and bovine ADAM10 were recently cloned and designated ADAM 10, kuzbanian (KUZ), and MADM, respectively. ADAM10 mRNA is expressed in a variety of human and bovine tissues.
Background References:
  • Rosendahl et al. J Biol Chem 1997;272:24588-93.
  • Pan and Rubin. Cell 1997;90:271-80.
  • Qi et al. Science 1999;283:91-4.
  • Howard et al. Biochem J 1996;317:45-50. (RD1299).
Buffer:
ADAM10 Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Conjugate:
Unconjugated
DISCLAIMER:
Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.
Homology:
Predicted species reactivity based on immunogen sequence: Rat: (100%), Bovine: (100%), Mouse: (100%)
Immunogen:
ADAM10 antibody was raised against a 17 amino acid peptide near the carboxy terminus of human ADAM10.

The immunogen is located within the last 50 amino acids of ADAM10.
Isoforms:
Human ADAM10 has 2 isoforms, including isoform 1 (748aa, 84kD) and isoform 2 (447aa, 49kD). Mouse ADAM10 has one isoform (749aa, 84kD) and Rat ADAM10 also has one isoform (749aa, 84kD). ADAM10 of human, mouse and rat has precursor form (94kD) and mature form (68kD). 2051 can detect both precursor ADAM10 and mature ADAM10 in human, mouse and rat.
NCBI Gene ID #:
102
NCBI Official Name:
ADAM metallopeptidase domain 10
NCBI Official Symbol:
ADAM10
NCBI Organism:
Homo sapiens
Physical State:
Liquid
PREDICTED MOLECULAR WEIGHT:
85 kDa
Protein Accession #:
NP_001101
Protein GI Number:
4557251
Purification:
ADAM10 Antibody is Protein A purified.
Research Area:
Apoptosis,Cancer
Swissprot #:
O14672
User NOte:
Optimal dilutions for each application to be determined by the researcher.
Validation:

KO validation (Figure 1 and 2): Anti-ADAM10 antibody (2051) specificity was further verified by ADAM10 specific knockout. ADAM10 signal in was detected by 2051 in wild type of MEF cells and 293 cells, but not in ADAM10 knockout cells.

Independent Antibody Validation in Cell lines (Figure 3) shows similar ADAM10 expression profile in human and mouse cell lines detected by two independent anti-ADAM10 antibodies that recognize different epitopes, 2051 against C-terminus domain and 24-024 against human recombinant fusion protein.  ADAM10 proteins are detected in the most tested cell lines at different expression levels by the two independent antibodies. 

Regulated expression validation (Figure 12 and 14): ADAM10 expression detected by anit-ADAM10 antibodies (2051) was down-regulated when exposed to oxygen-glucose deprivation (OGD) or glutamate (Fig 8), and up-regulated in membrane compartment whereas down-regulated in intracellular fractions when treated with donepezil (Fig 14).

KD validation (Figure 15): Anti-ADAM10 antibody (2051) specificity was further verified by ADAM10 specific knockdown. ADAM10 signal in HEK/APP cells transfected with ADAM10 siRNA or Ferrochelatase siRNA (F) or N-methylprotoporphyrin IX siRNA (N) was disrupted in comparison with that in mock-transfected cells (M) or cells transfected with TACE siRNA (T).

References

  • Al-Tamimi et al. Pathologic shear triggers shedding of vascular receptors: a novel mechanism for down-regulation of platelet glycoprotein VI in stenosed coronary vessels. Blood. 2012;119(18):4311-20. PMID: 22431567
  • Hurtado et al. TACE/ADAM17-TNF-alpha pathway in rat cortical cultures after exposure to oxygen-glucose deprivation or glutamate. J Cereb Blood Flow Metab. 2002;22(5):576-85.PMID: 11973430
  • Coma et al. Oxidative stress triggers the amyloidogenic pathway in human vascular smooth muscle cells. Neurobiol Aging. 2008;29(7):969-80. PMID: 17306421
  • Mosser et al. The adipocyte differentiation protein APMAP is an endogenous suppressor of A? production in the brain. Hum Mol Genet. 2015;24(2):371-82. PMID: 25180020
  • Zimmermann et al. Acetylcholinesterase inhibitors increase ADAM10 activity by promoting its trafficking in neuroblastoma cell lines. J Neurochem. 2004;90(6):1489-99.PMID: 15341532
  • Gatta et al. Inhibition of heme synthesis alters Amyloid Precursor Protein processing. J Neural Transm (Vienna). 2009;116(1):79-88. PMID: 19002554
  • Zimmermann et al. Cholinesterase inhibitors influence APP metabolism in Alzheimer disease patients. Neurobiol Dis. 2005;19(1-2):237-42. PMID: 15837579
  • Kabacik and Raj. Ionising radiation increases permeability of endothelium through ADAM10-mediated cleavage of VE-cadherin. Oncotarget. 2017;8(47):82049-82063. PMID: 29137243
  • Colciaghi et al. [alpha]-Secretase ADAM10 as well as [alpha]APPs is reduced in platelets and CSF of Alzheimer disease patients. Mol Med. 2002;8(2):67-74. PMID: 12080182
  • Bandyopadhyay et al. Interleukin-1alpha stimulates non-amyloidogenic pathway by alpha-secretase(ADAM-10 and ADAM-17) cleavage of APP in human astrocytic cells involving p38 MAP kinase. J Neurosci Res. 2006;84(1):106-18. PMID: 16724341
  • Hiltunen et al. Ubiquilin 1 modulates amyloid precursor protein trafficking and Abeta secretion. J Biol Chem. 2006;281(43):32240-53. PMID: 16945923
  • Harjes et al. Fatty acid-binding protein 4, a point of convergence for angiogenic and metabolic signaling pathways in endothelial cells. J Biol Chem. 2014;289(33):23168-76. PMID: 24939870
  • Levin-Allerhand et al. 17Alpha-estradiol and 17beta-estradiol treatments are effective in lowering cerebral amyloid-beta levels in AbetaPPSWE transgenic mice. J Alzheimers Dis. 2002;4(6):449-57. PMID: 12515896
  • Peng et al. Huperzine A regulates amyloid precursor protein processing via protein kinase C and mitogen-activated protein kinase pathways in neuroblastoma SK-N-SH cells over-expressing wild type human amyloid precursor protein 695. Neuroscience. 2007;150(2):386-95. PMID: 17945434
  • Smith et al. Compositions and methods for inhibiting beta amyloid secretion. U.S. Patent application publication. 2013. Pub. No.: US 2013/0158112 A1.
  • Can Zhang. Identifying Genes That Encode APP Metabolism Modulators On Chromosome 9q22. Drexel University, Philadelphia, PA. PhD Thesis, 2007.

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