| Applications: |
CLIA |
| Reactivity: |
Human |
| Note: |
FOR SCIENTIFIC EDUCATIONAL RESEARCH USE ONLY (RUO). MUST NOT BE USED IN DIAGNOSTIC OR OTHER MEDICAL APPLICATIONS. |
| Sensitivity: |
0.09ng/mL |
| Detection Limit: |
0.16~10ng/mL |
| Short Description: |
This human SMAD3 kit is a highly sensitive in-vitro chemiluminescent immunoassay for the measurement of trace amounts of analytes. |
| Storage Instruction: |
If unopened the kit may be stored at 2-8°C for up to 1 month. If the kit will not be used within 1 month, store the components separately, according to the component table in the manual. |
| Assay Time: |
3.5h |
| Detection: |
Chemiluminescence |
| Gene Symbol: |
SMAD3 |
| Gene ID: |
4088 |
| Uniprot ID: |
SMAD3_HUMAN |
| Specificity: |
This kit recognizes Human Smad3/MADH3 in samples. No significant cross-reactivity or interference between Human Smad3/MADH3 and analogues was observed. |
| Sample Type: |
Serum, plasma and other biological fluids |
| Post Translational Modifications | Phosphorylated on serine and threonine residues. Enhanced phosphorylation in the linker region on Thr-179, Ser-204 and Ser-208 on EGF and TGF-beta treatment. Ser-208 is the main site of MAPK-mediated phosphorylation. CDK-mediated phosphorylation occurs in a cell-cycle dependent manner and inhibits both the transcriptional activity and antiproliferative functions of SMAD3. This phosphorylation is inhibited by flavopiridol. Maximum phosphorylation at the G(1)/S junction. Also phosphorylated on serine residues in the C-terminal SXS motif by TGFBR1 and ACVR1. TGFBR1-mediated phosphorylation at these C-terminal sites is required for interaction with SMAD4, nuclear location and transactivational activity, and appears to be a prerequisite for the TGF-beta mediated phosphorylation in the linker region. Dephosphorylated in the C-terminal SXS motif by PPM1A. This dephosphorylation disrupts the interaction with SMAD4, promotes nuclear export and terminates TGF-beta-mediated signaling. Phosphorylation at Ser-418 by CSNK1G2/CK1 promotes ligand-dependent ubiquitination and subsequent proteasome degradation, thus inhibiting SMAD3-mediated TGF-beta responses. Phosphorylated by PDPK1. Acetylation in the nucleus by EP300 in the MH2 domain regulates positively its transcriptional activity and is enhanced by TGF-beta. Poly-ADP-ribosylated by PARP1 and PARP2. ADP-ribosylation negatively regulates SMAD3 transcriptional responses during the course of TGF-beta signaling. Ubiquitinated. Monoubiquitinated, leading to prevent DNA-binding. Deubiquitination by USP15 alleviates inhibition and promotes activation of TGF-beta target genes. Ubiquitinated by RNF111, leading to its degradation: only SMAD3 proteins that are 'in use' are targeted by RNF111, RNF111 playing a key role in activating SMAD3 and regulating its turnover. Undergoes STUB1-mediated ubiquitination and degradation. |
| Function | Receptor-regulated SMAD (R-SMAD) that is an intracellular signal transducer and transcriptional modulator activated by TGF-beta (transforming growth factor) and activin type 1 receptor kinases. Binds the TRE element in the promoter region of many genes that are regulated by TGF-beta and, on formation of the SMAD3/SMAD4 complex, activates transcription. Also can form a SMAD3/SMAD4/JUN/FOS complex at the AP-1/SMAD site to regulate TGF-beta-mediated transcription. Has an inhibitory effect on wound healing probably by modulating both growth and migration of primary keratinocytes and by altering the TGF-mediated chemotaxis of monocytes. This effect on wound healing appears to be hormone-sensitive. Regulator of chondrogenesis and osteogenesis and inhibits early healing of bone fractures. Positively regulates PDPK1 kinase activity by stimulating its dissociation from the 14-3-3 protein YWHAQ which acts as a negative regulator. |
| Protein Name | Mothers Against Decapentaplegic Homolog 3Mad Homolog 3Mad3Mothers Against Dpp Homolog 3Hmad-3Jv15-2Smad Family Member 3Smad 3Smad3Hsmad3 |
| Database Links | Reactome: R-HSA-1181150Reactome: R-HSA-1502540Reactome: R-HSA-2173788Reactome: R-HSA-2173789Reactome: R-HSA-2173795Reactome: R-HSA-2173796Reactome: R-HSA-3304356Reactome: R-HSA-3311021Reactome: R-HSA-3315487Reactome: R-HSA-3656532Reactome: R-HSA-5689880Reactome: R-HSA-8941855Reactome: R-HSA-8952158Reactome: R-HSA-9008059Reactome: R-HSA-9013695Reactome: R-HSA-9615017Reactome: R-HSA-9617828Reactome: R-HSA-9735871Reactome: R-HSA-9754189Reactome: R-HSA-9796292Reactome: R-HSA-9823730 |
| Cellular Localisation | CytoplasmNucleusCytoplasmic And Nuclear In The Absence Of Tgf-BetaOn Tgf-Beta StimulationMigrates To The Nucleus When Complexed With Smad4Through The Action Of The Phosphatase Ppm1aReleased From The Smad2/Smad4 ComplexAnd Exported Out Of The Nucleus By Interaction With Ranbp1Co-Localizes With Lemd3 At The Nucleus Inner MembraneMapk-Mediated Phosphorylation Appears To Have No Effect On Nuclear ImportPdpk1 Prevents Its Nuclear Translocation In Response To Tgf-BetaLocalized Mainly To The Nucleus In The Early Stages Of Embryo Development With Expression Becoming Evident In The Cytoplasm Of The Inner Cell Mass At The Blastocyst Stage |
| Alternative CLIA Names | Mothers Against Decapentaplegic Homolog 3 CLIA kitMad Homolog 3 CLIA kitMad3 CLIA kitMothers Against Dpp Homolog 3 CLIA kitHmad-3 CLIA kitJv15-2 CLIA kitSmad Family Member 3 CLIA kitSmad 3 CLIA kitSmad3 CLIA kitHsmad3 CLIA kitSMAD3 CLIA kitMADH3 CLIA kit |
| Specificity | This kit recognizes Human Smad3/MADH3 in samples. No significant cross-reactivity or interference between Human Smad3/MADH3 and analogues was observed. |
| Reproducibility | Both intra-CV and inter-CV are |
Information sourced from Uniprot.org
| Item | Specifications | Storage |
| Micro CLIA Plate (Dismountable) | 96T: 8 wells ×12 strips strips | -20℃, 6 months |
| Reference Standard | 96T: 2 vials 48T: 1 vial | -20℃, 6 months |
| Concentrated Biotinylated Detection Ab (100×) | 96T: 1 vial, 120 μL 60 μL | -20℃, 6 months |
| Concentrated HRP Conjugate (100×) | 96T: 1 vial, 120 μL 60 μL | -20℃ (Protect from light), 6 months |
| Reference Standard & Sample Diluent | 1 vial, 20 mL | 2-8°C, 6 months |
| Biotinylated Detection Ab Diluent | 1 vial, 14 mL | 2-8°C, 6 months |
| HRP Conjugate Diluent | 1 vial, 14 mL | 2-8°C, 6 months |
| Concentrated Wash Buffer (25×) | 1 vial, 30 mL | 2-8°C, 6 months |
| Substrate Reagent A | 1 vial, 5 mL | 2-8℃ (Protect from light) |
| Substrate Reagent B | 1 vial, 5 mL | 2-8℃ (Protect from light) |
| Plate Sealer | 5 pieces | |
| Manual | 1 copy | |
| Certificate of Analysis | 1 copy | |
| Sample Type | Range (%) | Average Recovery (%) |
| Serum(n=8) | 94-109 | 101 |
| EDTA plasma(n=8) | 101-113 | 107 |
| Cell culture media(n=8) | 87-101 | 94 |
| | Intra-assay Precision | Intra-assay Precision | Intra-assay Precision | Inter-assay Precision | Inter-assay Precision | Inter-assay Precision |
| Sample | 1.00 | 2.00 | 3.00 | 1.00 | 2.00 | 3.00 |
| n | 20.00 | 20.00 | 20.00 | 20.00 | 20.00 | 20.00 |
| Mean (ng/mL) | 0.54 | 1.01 | 3.87 | 0.56 | 1.06 | 4.07 |
| Standard deviation | 0.06 | 0.11 | 0.38 | 0.06 | 0.10 | 0.42 |
| CV (%) | 11.11 | 10.89 | 9.82 | 10.71 | 9.43 | 10.32 |
12 months for antibodies. 6 months for ELISA Kits. Please see website T&Cs for further guidance
