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Catalog No: OKCD02867
Size:96WELLS
Price: $725.00
SKU
OKCD02867
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Datasheets/ManualsPrintable datasheet for Smad4 ELISA kit (Rat)
Product Info
Predicted Species ReactivityRat
ApplicationEnzyme-linked Immunosorbent assay-Sandwich
ELISA Kit Detection MethodColormetric|Regular detection
ELISA Kit Duration3h
ELISA Kit PrincipleThe test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Mothers Against Decapentaplegic Homolog 4 (Smad4). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Mothers Against Decapentaplegic Homolog 4 (Smad4). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Mothers Against Decapentaplegic Homolog 4 (Smad4), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm +/- 10nm. The concentration of Mothers Against Decapentaplegic Homolog 4 (Smad4) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
ELISA Kit Range0.156-10ng/mL
ELISA Kit ReproducibilityIntra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Mothers Against Decapentaplegic Homolog 4 (Smad4) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Mothers Against Decapentaplegic Homolog 4 (Smad4) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%
ELISA Kit Component
ComponentAmount
Anti-SMAD4 Microplate96 Wells (12 x 8 Well strips)
SMAD4 Lyophilized Standard2
100X Biotinylated SMAD4 Detector Antibody1 x 120 uL
100X Avidin-HRP Conjugate1 x 120 uL
Standard Diluent1 x 20 mL
Detector Antibody Diluent1 x 12 mL
Conjugate Diluent1 x 12 mL
30X Wash Buffer1 x 20 mL
Stop Solution1 x 6 mL
TMB Substrate1 x 9 mL
Reconstitution and Storage2°C to 8°C, -20°C
Sample TypeTissue homogenates, cell lysates and other biological fluids
Sensitivity0.054 ng/mL
SpecificityThis assay has high sensitivity and excellent specificity for detection of Mothers Against Decapentaplegic Homolog 4 (Smad4). No significant cross-reactivity or interference between Mothers Against Decapentaplegic Homolog 4 (Smad4) and analogues was observed.
Assay InfoAssay Methodology: Quantitative Sandwich ELISA
Protocol Information
Gene SymbolSMAD4
Gene Full NameSMAD family member 4
Alias SymbolsMadh4
NCBI Gene Id50554
Protein NameMothers against decapentaplegic homolog 4
Description of TargetCommon SMAD (co-SMAD) is the coactivator and mediator of signal transduction by TGF-beta (transforming growth factor). Component of the heterotrimeric SMAD2/SMAD3-SMAD4 complex that forms in the nucleus and is required for the TGF-mediated signaling. Promotes binding of the SMAD2/SMAD4/FAST-1 complex to DNA and provides an activation function required for SMAD1 or SMAD2 to stimulate transcription. Component of the multimeric SMAD3/SMAD4/JUN/FOS complex which forms at the AP1 promoter site; required for synergistic transcriptional activity in response to TGF-beta. Acts synergistically with SMAD1 and YY1 in bone morphogenetic protein (BMP)-mediated cardiac-specific gene expression. Binds to SMAD binding elements (SBEs) (5'-GTCT/AGAC-3') within BMP response element (BMPRE) of cardiac activating regions. May act as a tumor suppressor. Positively regulates PDPK1 kinase activity by stimulating its dissociation from the 14-3-3 protein YWHAQ which acts as a negative regulator. In muscle physiology, plays a central role in the balance between atrophy and hypertrophy. When recruited by MSTN, promotes atrophy response via phosphorylated SMAD2/4. MSTN decrease causes SMAD4 release and subsequent recruitment by the BMP pathway to promote hypertrophy via phosphorylated SMAD1/5/8.
Uniprot IDO70437
Protein Accession #NP_062148.1
Nucleotide Accession #NM_019275.3
Protein Size (# AA)552
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