TWEAK is a type II transmembrane protein belonging to the TNF superfamily (1). It contains a short cytoplasmic domain (amino acids (aa) 1-18), the transmembrane domain (aa 19-42) and an extracellular domain (aa 43-249). The extracellular domains of human and murine TWEAK share 89% aa sequence identity. A soluble form of TWEAK is generated from the membrane-associated molecules by proteolytic cleavage after Arg 93 suggesting that TWEAK may have long-range effects. TWEAK is expressed widely in many tissues and cells. At least two receptors that bind TWEAK have been identified (2-4). Death Receptor 3 (DR3), also known as TNFRSF12, Apo-3, LARD, WSL-1, or TRAMP, is a TNF receptor superfamily member that is expressed predominantly in tissues with high lymphocyte content (2). It has been suggested that induction of cell death by TWEAK-DR3 interaction involves the activation of NF-kappa B. In cells that lack DR3, alternate pathways of TWEAK-induced cell death mediated by receptors distinct from DR3 have been suggested (5, 6). TWEAK receptor (TWEAKR, alternatively known as FN14), is a novel TNF receptor superfamily member that also binds TWEAK (3, 4). It is a mitogen-inducible gene that is expressed in fibroblasts, hepetocellular carcinomas and endothelial cells. TWEAK-TWEAKR interaction has been shown to play a role in endothelial cell growth and migration. This effect of TWEAK is not mediated by an up-regulation of VEGF (7).
Key Product Details
Species Reactivity
Validated:
Human
Cited:
Human, Mouse
Applications
Validated:
Western Blot, Neutralization, Immunocytochemistry
Cited:
Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Neutralization
Label
Unconjugated
Antibody Source
Polyclonal Goat IgG
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Product Specifications
Immunogen
E. coli-derived recombinant human TWEAK
Arg93-His249
Accession # Q4ACW9
Arg93-His249
Accession # Q4ACW9
Specificity
Detects human TWEAK in direct ELISAs and Western blots. In direct ELISAs, less than 5% cross-reactivity with recombinant human (rh) BAFF, rhFas Ligand, rhGITR Ligand, rhOX40 Ligand, rhTRAIL and less than 1% cross-reactivity with rhAPRIL, rhLIGHT, rhTNF-alpha, rhVEGI, and rhTRANCE is observed.
Clonality
Polyclonal
Host
Goat
Isotype
IgG
Endotoxin Level
<0.10 EU per 1 μg of the antibody by the LAL method.
Scientific Data Images for Human TWEAK/TNFSF12 Antibody
TWEAK/TNFSF12 Inhibition of Cell Proliferation and Neutralization by Human TWEAK/TNFSF12 Antibody.
Recombinant Human TWEAK/TNFSF12 (Catalog # 1090-TW) inhibits proliferation in the the HT-29 human colon adenocarcinoma cell line in a dose-dependent manner (orange line). Activity elicited by Recombinant Human TWEAK/TNFSF12 (1 µg/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human TWEAK/ TNFSF12 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1090). The ND50 is typically 2-8 µg/mL.TWEAK/TNFSF12 in MDA-MB-231 Human Cell Line.
TWEAK/TNFSF12 was detected in immersion fixed MDA-MB-231 human breast cancer cell line using 10 µg/mL Goat Anti-Human TWEAK/TNFSF12 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1090) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.Applications for Human TWEAK/TNFSF12 Antibody
Application
Recommended Usage
Immunocytochemistry
5-15 µg/mL
Sample: Immersion fixed MDA-MB-231 human breast cancer cell line
Sample: Immersion fixed MDA-MB-231 human breast cancer cell line
Western Blot
0.1 µg/mL
Sample: Recombinant Human TWEAK/TNFSF12 (Catalog # 1090-TW)
Sample: Recombinant Human TWEAK/TNFSF12 (Catalog # 1090-TW)
Neutralization
Measured by its ability to neutralize TWEAK/TNFSF12-induced inhibition of proliferation in the HT‑29 human colon adenocarcinoma cell line. Yu, K. Y. et al. (1999) J. Biol. Chem. 274:13733; Harrop, J. A. et al. (1998) J. Biol. Chem. 273:27548The Neutralization Dose (ND50) is typically 2-8 µg/mL in the presence of 1 µg/mL Recombinant Human TWEAK/TNFSF12.
Reviewed Applications
Read 1 review rated 5 using AF1090 in the following applications:
Formulation, Preparation, and Storage
Purification
Antigen Affinity-purified
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
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Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Calculators
Background: TWEAK/TNFSF12
References
- Chicheportiche, Y. et al. (1997) J. Biol Chem. 272:32401.
- Marsters, S. et al. (1998) Current Biol. 8:525.
- Wiley, S.R. et al. (2001) Immunity, 15:837.
- Feng, S-L.Y. et al. (2000) Am J. Path. 156:1253.
- Nakayama, M. et al. (2002) J. Immunol. 168:734.
- Schneider, P. et al. (1999) Eur. J. Immunol., 29:1785.
- Lynch, C. et al. (1998) J. Biol. Chem. 274:8455.
Long Name
TNF-like and Weak Inducer of Apoptosis
Alternate Names
TNFSF12
Gene Symbol
TNFSF12
UniProt
Additional TWEAK/TNFSF12 Products
Product Documents for Human TWEAK/TNFSF12 Antibody
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human TWEAK/TNFSF12 Antibody
For research use only
Related Research Areas
Citations for Human TWEAK/TNFSF12 Antibody
Customer Reviews for Human TWEAK/TNFSF12 Antibody (1)
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Application: ELISASample Tested: Serum and PlasmaSpecies: HumanVerified Customer | Posted 11/08/2019This antibody was used as the detection in an ELISA with MAB1090 as the capture.
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- ClariTSA™ Fluorophore Kits
- Detection & Visualization of Antibody Binding
- ICC Cell Smear Protocol for Suspension Cells
- ICC Immunocytochemistry Protocol Videos
- ICC for Adherent Cells
- Immunocytochemistry (ICC) Protocol
- Immunocytochemistry Troubleshooting
- Immunofluorescence of Organoids Embedded in Cultrex Basement Membrane Extract
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Fluorescent ICC Staining of Cell Smears - Graphic
- Protocol for the Fluorescent ICC Staining of Cultured Cells on Coverslips - Graphic
- Protocol for the Preparation and Fluorescent ICC Staining of Cells on Coverslips
- Protocol for the Preparation and Fluorescent ICC Staining of Non-adherent Cells
- Protocol for the Preparation and Fluorescent ICC Staining of Stem Cells on Coverslips
- Protocol for the Preparation of a Cell Smear for Non-adherent Cell ICC - Graphic
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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