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<strong>Figure 1 Western Blot Validation of TACE in Human Cell Lines</strong><br> Loading: 15 μg of lysates per lane. Antibodies: TACE (1 µg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lanes: HeLa (A,D), Jurkat (B, E), Raji (C,F) in the absence (A-C) or presence (E-F) of blocking peptide.
<strong>Figure 2 KO Validation in HeLa Cells</strong><br> Loading: 10 μg of HeLa WT cell lysates or TACE KO cell lysates. Antibodies: TACE 1131 (0.25 μg/mL) and beta-actin 3779 (1 μg/mL), 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 3 Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines</strong><br> Loading: 15 μg of lysates per lane. Antibodies: TACE 1131 (0.5 μg/mL), TACE 22-001 (2 μg/mL), and GAPDH (0.02 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 4 Immunofluorescence Validation of TACE in HeLa Cells</strong><br>Immunofluorescent analysis of 4% paraformaldehyde-fixed HeLa cells labeling TACE with 1131 at 10 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green).
<strong>Figure 5 Immunocytochemistry Validation of TACE in HeLa Cells</strong><br>Immunohistochemical analysis of HeLa cells using anti-TACE antibody (1131) at 10 μg/ml. Cells was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 6 KD Validation of TACE in Monkey COS Cells. (Wang et al., 2006) </strong><br>COS cells stably expressing Pref-1A were transfected with control siRNA or TACE siRNA. TACE was detected in lysates by using the anti-TACE antibody (1131). TACE expression levels were markedly reduced in TACE knockdown cell lysate.
<strong>Figure 7 KD Validation of TACE in MDA-MB-435 Cells. (McGowan et al., 2007) </strong><br>ADAM-17 protein expression, following transfection with ADAM-17 shRNA (two clones) or neomycin-resistant negative control vector, was examined by immunoblot analysis with anti-ADAM-17 antibodies (1131).
<strong>Figure 8 Overexpression Validation of TACE in MCF-7 Cells. (McGowan et al., 2007) </strong><br>ADAM-17 (TACE) protein expression, following transfection of vector and ADAM-17 cDNA, was examined by immunoblot analysis with anti-ADAM-17 (1131) antibodies in MCF-7 cells. Increased ADAM-17 was detected in ADAM-17 transfected cells.
<strong>Figure 9 Induced Expression Validation of TACE in Rat Cortical Neurons (Hurtado et al., 2002) </strong><br>Effect of oxygen–glucose deprivation(OGD) or glutamate on the levels of TACE/ADAM17 in rat cortical cultures. Western blot analysis of TACE in homogenates from control, glutamate, and OGD-exposed cultures from a representative experiment.
<strong>Figure 10 Immunofluorescence Validation of TACE in Rat Cortical Neurons (Hurtado et al., 2002) </strong><br> Double immunostaining of control and glutamate-exposed rat cortical cultures. (A) Control cultures show TACE immunoreactivity at the cellular membrane of some microglial cells (B) Glutamate-exposed cultures show that most microglial cells express TACE immunoreactivity.(C) Control cultures show that TACE immunostaining does not colocalize with astrocytes [glial fibrillary acidic protein (GFAP)-positive cells]. (D) Astrocyte (GFAP-positive cell) showing TACE immunoreactivity in its surface after treatment with glutamate.
<strong>Figure 11 Immunofluorescence Validation of TACE in Rat Brain (Pradillo et al, 2005) </strong><br> Cellular localization of TACE. Double immunofluorescence staining of brain sections from sham-operated (SHAM; A, C, E) and IPC-exposed animals (IPC; B, D, F) of TACE (red) and the cellular markers (green) NeuN (neurons; A, B), GFAP (astrocytes; C, D) and L. esculentum lectin (microglia and endothelium; E, F). White arrows indicate TACE-positive cells.
Figure 1 Western Blot Validation of TACE in Human Cell Lines
Loading: 15 μg of lysates per lane. Antibodies: TACE (1 µg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lanes: HeLa (A,D), Jurkat (B, E), Raji (C,F) in the absence (A-C) or presence (E-F) of blocking peptide.
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TACE Antibody

CATALOG NUMBER: 1131

Clonality:
Polyclonal
Tested Applications:
ELISA, ICC, IF, WB
Host Species:
Rabbit
Species Reactivity:
Human, Rat
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human, Rat
Homology:
Predicted species reactivity based on immunogen sequence: mouse (94.1%)
Immunogen:
Anti-TACE antibody (1131) was raised against a peptide corresponding to 17 amino acids near the carboxy terminus of human TACE.
The immunogen is located within the last 50 amino acids of TACE.
Conjugate:
Unconjugated
Tested Applications:
ELISA, ICC, IF, WB
Application Note:
WB: 0.25-2 μg/mL; ICC: 10 μg/mL; IF: 10 μg/mL.
Antibody validated: Western Blot in human and rat samples; Immunocytochemistry in human samples; and Immunofluorescence in human and rat samples. All other applications and species not yet tested.
Specificity:
80 to 130 kDa bands can be detected, which may represent mature protein, precursor, and glycosylated TACE.
Positive Control 1:
Cat. No. 1201 - HeLa Cell Lysate
Positive Control 2:
Cat. No. 1205 - Jurkat Cell Lysate
Positive Control 3:
Cat. No. 1207 - Raji Cell Lysate
Positive Control 4:
Cat. No. 17-001 - HeLa Cell Slide
Predicted Molecular Weight:
Predicted: 93kD
Observed: 93-125kD (Post-modification: 9 N-linked glycosylation)
Validation:

KO Validation (Figure 2): Anti-TACE antibodies (1131) specificity was further verified by TACE specific knockout. TACE signal was not detected in TACE knockout HeLa cells as compared to that in control wild type cells.

Independent Antibody Validation in Cell lines (Figure 3) shows similar TACE expression profile in human cell lines detected by two independent anti-TACE antibodies that recognize different epitopes, 1131 against C-terminus domain and  22-001 against a synthetic peptide to a sequence within amino acids 700-824 of human TACE.  TACE proteins are detected in most of the tested cell lines except 293 and A549 cell lines at different expression levels by the two independent antibodies.  

KD validation (Figure 6,7): Anti-TACE antibody (1131) specificity was verified by TACE TACE specific siRNA or shRNA knockdown. TACE signal in COS cells (Figure 6) and MDA-MB-435 cells (Figure 7) with TACE knockdown was disrupted in comparison with control.

Overexpression validation (Figure 8): Anti-TACE antibody (1131) detected high expression levels of TACE in MCF-7 cells transfected with TACE as compared to vector transfected cells.

Isoforms:
Human TACE has 2 isoforms, including isoform A (824aa, 93kD), isoform B (694aa, 78.5kD). This antibody detects only isoform A. Mouse TACE has 2 isoforms, long isoform (827aa, 93.1kD) and short isoform (655aa, 73.9kD). Rat TACE has only one isoform identified so far (827aa, 93kD).
Purification:
TACE Antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgG
physical-state:
Liquid
Buffer:
TACE Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
TACE antibody can be stored at 4°C for three months and -20°C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.
Ncbi Official Symbol:
ADAM17
Additional Names:
TACE Antibody: CSVP, TACE, NISBD, ADAM18, CD156B, CSVP, Disintegrin and metalloproteinase domain-containing protein 17, Snake venom-like protease, ADAM 17
Protein Accession Number:
NP_003174
Protein Gi Number:
73747889
Ncbi Gene Id Number:
6868
User Note:
Optimal dilutions for each application to be determined by the researcher.
Background:
TACE Antibody: Tumor-necrosis factor-alpha is a proinflammatory cytokine and contributes to a variety of inflammatory disease responses and programmed cell death. TNF-α is synthesized as a 26K type II membrane-bound precursor that is cleaved by a convertase to generate secreted 17K mature TNF-α. TNF-α converting enzyme (TACE) protein was recently purified and the human and mouse TACE cDNAs were cloned by several groups separately. TACE is a membrane-bound metalloprotease-disintegrin in the family of mammalian ADAM (for a disintegrin and metalloprotease). TACE also processes other cell surface proteins, including TNF receptor, TGFalpha, the L-selectin adhesion molecule, and alpha-cleavage of amyloid protein precursor (APP). TACE mRNA is expressed in a variety of human and murine tissues. TACE was selected as one of the few targets in cytokine activation by the Eighth International Conference of the Inflammation Research Association.
Background Reference 1:
Black et al. Nature 1997;385:729-733
Background Reference 2:
Moss et al. Nature 1997;385:733-736
Background Reference 3:
Mizui et al. Gene 1999;233:67-74
Background Reference 4:
Peschon et al. Science 1998;282:1281-4
Citation 1:
Wang et al. Ectodomain shedding of preadipocyte factor 1 (Pref-1) by tumor necrosis factor alpha converting enzyme (TACE) and inhibition of adipocyte differentiation. Mol Cell Biol. 2006 ;26(14):5421-35. PMID: 16809777
Citation 2:
McGowan et al. ADAM-17 expression in breast cancer correlates with variables of tumor progression. Clin Cancer Res. 2007;13(8):2335-43. PMID: 17438092
Citation 3:
Hurtado et al. TACE/ADAM17-TNF-alpha pathway in rat cortical cultures after exposure to oxygen-glucose deprivation or glutamate. J Cereb Blood Flow Meta. 2002 ;22(5):576-85. PMID: 11973430
Citation 4:
Pradillo et al. TNFR1 upregulation mediates tolerance after brain ischemic preconditioning. J Cereb Blood Flow Metab. 2005;25(2):193-203.PMID: 15647744
Citation 5:
McGowan et al. ADAM-17 predicts adverse outcome in patients with breast cancer. Ann Oncol. 2008;19(6):1075-81. PMID: 18238782
Citation 6:
García-Bueno et al. Peroxisome proliferator-activated receptor gamma activation decreases neuroinflammation in brain after stress in rats. Biol Psychiatry. 2005;57(8):885-94. PMID: 15820709
Citation 7:
Romera et al. In vitro ischemic tolerance involves upregulation of glutamate transport partly mediated by the TACE/ADAM17-tumor necrosis factor-alpha pathway. J Neurosci. 2004;24(6):1350-7. PMID: 14960606
Citation 8:
Zimmermann et al. Acetylcholinesterase inhibitors increase ADAM10 activity by promoting its trafficking in neuroblastoma cell lines. J Neurochem. 2004;90(6):1489-99PMID: 15341532
Citation 9:
Ge et al. Soluble TNF Regulates TACE via AP-2α Transcription Factor in Mouse Dendritic Cells. J Immunol. 2017;198(1):417-427PMID: 27852742
Citation 10:
Comabella et al. TNF-alpha converting enzyme (TACE) protein expression in different clinical subtypes of multiple sclerosis. J Neurol. 2006;253(6):701-6PMID: 16511637
Citation 11:
García-Bueno et al. The anti-inflammatory prostaglandin 15d-PGJ2 decreases oxidative/nitrosative mediators in brain after acute stress in rats. Psychopharmacology (Berl). 2005 ;180(3):513-22PMID: 15719215
Citation 12:
Gatta et l. Inhibition of heme synthesis alters Amyloid Precursor Protein processing. J Neural Transm (Vienna). 2009;116(1):79-88.PMID: 19002554
Citation 13:
Wu et al. ADAM-17 over-expression in gallbladder carcinoma correlates with poor prognosis of patients. Med Oncol. 2011;28(2):475-80PMID: 20300969
Citation 14:
Bigdeli et al. Preconditioning with sublethal ischemia or intermittent normobaric hyperoxia up-regulates glutamate transporters and tumor necrosis factor-alpha converting enzyme in the rat brain. J Stroke Cerebrovasc Dis. 2009;18(5):336-42. PMID: 19717015

FOR RESEARCH USE ONLY

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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.

CATALOG NUMBER:

1131

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

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