Control Lysate
(HeLa cells were grown in a T175 flask at 37 degree C, 5%CO2, in DMEM with 10% FBS and cultured to 95% confluence. The next day, cells were starved for 3 h. The medium was removed and cells were stimulated with 0.3 nM of TGF-betafor 60 min. The medium was removed and cells were lysed with 4 mL of 1X Lysis Buffer #2 containing 1 mM NaF and 2 mM Na3VO4, on an orbital shaker (400 rpm) for 30 min at room temperature. Soluble supernatants were collected after a 10 min centrifugation and tested using the two-plate assay protocol.)
SMAD2 assay kit
Phospho-SMAD2 (S465/S467) TR-FRET Cellular Assay Kit
Predicted: Mouse, Rat
Predicted: Mouse, Rat
Acc-phospho-SMAD2 (20 uL)
SMAD2 control lysate (100 uL)
5X Lysis Buffer #2 (1 mL)
10X TR-FRET Detection buffer (50 uL)
Control Lysate
(HeLa cells were grown in a T175 flask at 37 degree C, 5%CO2, in DMEM with 10% FBS and cultured to 95% confluence. The next day, cells were starved for 3 h. The medium was removed and cells were stimulated with 0.3 nM of TGF-betafor 60 min. The medium was removed and cells were lysed with 4 mL of 1X Lysis Buffer #2 containing 1 mM NaF and 2 mM Na3VO4, on an orbital shaker (400 rpm) for 30 min at room temperature. Soluble supernatants were collected after a 10 min centrifugation and tested using the two-plate assay protocol.)
Control Lysate
(HeLa cells were grown in a T175 flask at 37 degree C, 5%CO2, in DMEM with 10% FBS and cultured to 95% confluence. The next day, cells were starved for 3 h. The medium was removed and cells were stimulated with 0.3 nM of TGF-betafor 60 min. The medium was removed and cells were lysed with 4 mL of 1X Lysis Buffer #2 containing 1 mM NaF and 2 mM Na3VO4, on an orbital shaker (400 rpm) for 30 min at room temperature. Soluble supernatants were collected after a 10 min centrifugation and tested using the two-plate assay protocol.)
Testing Data
(HeLa cells were seeded at 80,000 cells/well and cultured overnight at 37 degree C, 5% CO2, in DMEM with 10% FBS. Cells were starved for 3 h then treated with either TGF-beta for 60 min (stimulation) or with SB 431542 for 30 min then 0.3 nM TGF-beta (inhibition). The media was removed from the wells, and cells were lysed with 50 uL/well of 1X Lysis Buffer #2 containing 1 mM NaF and 2 mM Na3VO4, on an orbital shaker (400 rpm) for 30 min at room temperature. The lysates were then analyzed for phospho-SMAD2 (S465/S467) using the two-plate assay protocol. The TR-FRET signal was recorded at 665 and 615 nm (EnVision lamp excitation) after a 2 h incubation period at room temperature.)
Testing Data
(HeLa cells were seeded at 80,000 cells/well and cultured overnight at 37 degree C, 5% CO2, in DMEM with 10% FBS. Cells were starved for 3 h then treated with either TGF-beta for 60 min (stimulation) or with SB 431542 for 30 min then 0.3 nM TGF-beta (inhibition). The media was removed from the wells, and cells were lysed with 50 uL/well of 1X Lysis Buffer #2 containing 1 mM NaF and 2 mM Na3VO4, on an orbital shaker (400 rpm) for 30 min at room temperature. The lysates were then analyzed for phospho-SMAD2 (S465/S467) using the two-plate assay protocol. The TR-FRET signal was recorded at 665 and 615 nm (EnVision lamp excitation) after a 2 h incubation period at room temperature.)
A superior alternative to ELISA
Features :: Benefits
• TR-FRET technology :: Proven, sensitive and robust assay technology
• Homogeneous, add-and-read assay :: No wash, no separation steps
• < 1 hour bench time :: Faster, easier than ELISA
• Adherent or suspension cells :: Higher flexibility
• Validated assays :: Results you can trust
• Small sample volume (15uL) :: Save precious samples
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• Signal stable over time :: Flexibility to read plate
• Kits available in 100 and 500 assay points :: Cost-effective and flexible
• Kits available for phospho, total and phospho+total protein :: Allows multiple protein measurements in one plate from one sample
NCBI and Uniprot Product Information
NCBI Description
The protein encoded by this gene belongs to the SMAD, a family of proteins similar to the gene products of the Drosophila gene 'mothers against decapentaplegic' (Mad) and the C. elegans gene Sma. SMAD proteins are signal transducers and transcriptional modulators that mediate multiple signaling pathways. This protein mediates the signal of the transforming growth factor (TGF)-beta, and thus regulates multiple cellular processes, such as cell proliferation, apoptosis, and differentiation. This protein is recruited to the TGF-beta receptors through its interaction with the SMAD anchor for receptor activation (SARA) protein. In response to TGF-beta signal, this protein is phosphorylated by the TGF-beta receptors. The phosphorylation induces the dissociation of this protein with SARA and the association with the family member SMAD4. The association with SMAD4 is important for the translocation of this protein into the nucleus, where it binds to target promoters and forms a transcription repressor complex with other cofactors. This protein can also be phosphorylated by activin type 1 receptor kinase, and mediates the signal from the activin. Alternatively spliced transcript variants have been observed for this gene. [provided by RefSeq, May 2012]
Uniprot Description
SMAD2: ubiquitously expressed transcription factor phosphorylated and activated by TGF-beta receptor-type kinases. Participates in a wide range of critical processes including morphogenesis, cell-fate determination, proliferation, differentiation and apoptosis. Phosphorylated Smads dimerize with collaborating Smad4 and are translocated into the nucleus, where the transcription of target genes is stimulated. Two alternatively spliced isoforms have been described.
Protein type: DNA-binding; Transcription factor
Chromosomal Location of Human Ortholog: 18q21.1
Cellular Component: nucleoplasm; transcription factor complex; cytoplasm; nuclear chromatin; nucleus; cytosol
Molecular Function: transcription activator binding; metal ion binding; transcription factor binding; transforming growth factor beta receptor, pathway-specific cytoplasmic mediator activity; phosphatase binding; protein binding; DNA binding; ubiquitin protein ligase binding; double-stranded DNA binding; chromatin binding; SMAD binding; transcription factor activity; transforming growth factor beta receptor binding
Biological Process: developmental growth; somatic stem cell maintenance; positive regulation of transcription, DNA-dependent; activin receptor signaling pathway; paraxial mesoderm morphogenesis; gastrulation; primary microRNA processing; negative regulation of transcription from RNA polymerase II promoter; palate development; post-embryonic development; anterior/posterior pattern formation; negative regulation of cell proliferation; regulation of transforming growth factor beta receptor signaling pathway; ureteric bud development; transforming growth factor beta receptor signaling pathway; pancreas development; embryonic foregut morphogenesis; response to glucose stimulus; positive regulation of BMP signaling pathway; pericardium development; transcription initiation from RNA polymerase II promoter; cell fate commitment; regulation of binding; transcription, DNA-dependent; in utero embryonic development; common-partner SMAD protein phosphorylation; embryonic cranial skeleton morphogenesis; SMAD protein complex assembly; organ growth; zygotic determination of dorsal/ventral axis; mesoderm formation; endoderm formation; insulin secretion; gene expression; positive regulation of transcription from RNA polymerase II promoter; negative regulation of transcription, DNA-dependent; negative regulation of transforming growth factor beta receptor signaling pathway; lung development
Research Articles on SMAD2
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Product Notes
The Human Predicted: Mouse, Rat SMAD2 smad2 (Catalog #AAA298058) is an Assay Kit and is intended for research purposes only. The product is available for immediate purchase. The AAA298058 Assay Kit recognizes Human Predicted: Mouse, Rat SMAD2. AAA Biotech's SMAD2 can be used in a range of immunoassay formats including, but not limited to, Cell-Based Assays. Researchers should empirically determine the suitability of the SMAD2 smad2 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "SMAD2, Assay Kit" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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