Rat Signaling ELISA Kits 2
Rat BMP-4 (Bone Morphogenetic Protein 4) CLIA Kit (RTES00069)
- SKU:
- RTES00069
- Product Type:
- ELISA Kit
- ELISA Type:
- CLIA Kit
- Size:
- 96 Assays
- Sensitivity:
- 4.69pg/mL
- Range:
- 7.81-500pg/mL
- ELISA Type:
- Sandwich
- Synonyms:
- BMP2B, BMP2B1, MCOPS6, ZYME, DVR4, OFC11, ZYME
- Reactivity:
- Rat
- Sample Type:
- Serum, plasma and other biological fluids
- Research Area:
- Cell Biology
Description
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Rat |
| Detection method: | Chemiluminescence |
| Detection range: | 7.81-500 pg/mL |
| Sensitivity: | 4.69 pg/mL |
| Sample volume: | 100µL |
| Sample type: | Serum, plasma and other biological fluids |
| Repeatability: | CV < 15% |
| Specificity: | This kit recognizes Rat BMP-4 in samples. No significant cross-reactivity or interference between Rat BMP-4 and analogues was observed. |
This kit uses Sandwich-CLIA as the method. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Rat BMP-4. Standards or samples are added to the appropriate micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Rat BMP-4 and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Rat BMP-4, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured spectrophotometrically by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Rat BMP-4. The concentration of Rat BMP-4 in the samples can be calculated by comparing the RLU of the samples to the standard curve.
| UniProt Protein Function: | BMP4: Induces cartilage and bone formation. Also act in mesoderm induction, tooth development, limb formation and fracture repair. Acts in concert with PTHLH/PTHRP to stimulate ductal outgrowth during embryonic mammary development and to inhibit hair follicle induction. Homodimer; disulfide-linked. Interacts with GREM2. Part of a complex consisting of TWSG1 and CHRD. Interacts with the serine proteases, HTRA1 and HTRA3; the interaction with either inhibits BMP4-mediated signaling. The HTRA protease activity is required for this inhibition. Interacts with SOSTDC1. Expressed in the lung and lower levels seen in the kidney. Present also in normal and neoplastic prostate tissues, and prostate cancer cell lines. Belongs to the TGF-beta family. |
| UniProt Protein Details: | Protein type:Secreted; Secreted, signal peptide Cellular Component: extracellular space; proteinaceous extracellular matrix; cytoplasm; extracellular region Molecular Function:heparin binding; protein homodimerization activity; growth factor activity; cytokine activity; transforming growth factor beta receptor binding; chemoattractant activity Biological Process: negative regulation of MAP kinase activity; activation of MAPKK activity; positive regulation of apoptosis; positive regulation of transcription, DNA-dependent; embryonic skeletal development; negative regulation of chondrocyte differentiation; mesodermal cell differentiation; telencephalon regionalization; germ cell development; regulation of protein import into nucleus; BMP signaling pathway; anatomical structure formation; mesonephros development; kidney development; endochondral ossification; regulation of odontogenesis of dentine-containing teeth; embryonic limb morphogenesis; positive regulation of cardiac muscle fiber development; negative regulation of immature T cell proliferation in the thymus; tongue morphogenesis; cell fate commitment; camera-type eye development; regulation of smooth muscle cell proliferation; neuron fate commitment; response to testosterone stimulus; camera-type eye morphogenesis; regulation of gene expression; retina development in camera-type eye; response to mechanical stimulus; negative regulation of mitosis; positive regulation of epidermal cell differentiation; smooth muscle cell differentiation; positive regulation of transcription from RNA polymerase II promoter; embryonic digit morphogenesis; negative regulation of apoptosis; tissue development; wound healing; positive regulation of protein binding; cloacal septation; negative regulation of transcription from RNA polymerase II promoter; anatomical structure regression; negative regulation of cell proliferation; inner ear receptor cell differentiation; ureteric bud development; intermediate mesodermal cell differentiation; forebrain development; positive regulation of cell proliferation; angiogenesis; embryonic morphogenesis; positive regulation of BMP signaling pathway; common-partner SMAD protein phosphorylation; negative regulation of T cell differentiation in the thymus; positive regulation of bone mineralization; positive regulation of ossification; embryonic skeletal morphogenesis; odontogenesis of dentine-containing teeth; osteoblast differentiation; positive regulation of osteoblast differentiation; cell proliferation; blood vessel endothelial cell proliferation during sprouting angiogenesis; telencephalon development; ureteric bud branching; regulation of cell fate commitment; brain development; positive regulation of neuron differentiation; anterior/posterior axis specification; lung development; renal system process; macrophage differentiation; heart development; response to glucocorticoid stimulus; lymphoid progenitor cell differentiation; response to organic cyclic substance; post-embryonic development; positive regulation of endothelial cell differentiation; positive chemotaxis; induction of an organ; erythrocyte differentiation; chondrocyte differentiation; mesoderm development; specification of organ position; monocyte differentiation; embryonic cranial skeleton morphogenesis; negative regulation of striated muscle development; mesoderm formation; branching morphogenesis of a tube; negative regulation of phosphorylation; positive regulation of endothelial cell proliferation; hemopoietic progenitor cell differentiation; steroid hormone mediated signaling; negative regulation of transcription, DNA-dependent; positive regulation of cell differentiation; metanephros development; alveolus development; positive regulation of epithelial cell proliferation; positive regulation of smooth muscle cell proliferation; positive regulation of collagen biosynthetic process; embryonic hindlimb morphogenesis; response to estradiol stimulus; negative regulation of cell cycle; odontogenesis; smooth muscle development; vasculature development; ovarian follicle development; regulation of cell differentiation; regulation of smooth muscle cell differentiation; skeletal development; dorsoventral neural tube patterning; negative regulation of epithelial cell proliferation; blood vessel development; smoothened signaling pathway; response to retinoic acid; negative regulation of oligodendrocyte differentiation; eye development; pituitary gland development; neural tube closure; positive regulation of protein amino acid phosphorylation; negative regulation of myoblast differentiation; mesodermal cell fate determination; growth |
| NCBI Summary: | plays a role in induction of cell proliferation [RGD, Feb 2006] |
| UniProt Code: | Q06826 |
| NCBI GenInfo Identifier: | 543902 |
| NCBI Gene ID: | 25296 |
| NCBI Accession: | Q06826. 1 |
| UniProt Related Accession: | Q06826 |
| Molecular Weight: | 19. 5kDa |
| NCBI Full Name: | Bone morphogenetic protein 4 |
| NCBI Synonym Full Names: | bone morphogenetic protein 4 |
| NCBI Official Symbol: | Bmp4 |
| NCBI Official Synonym Symbols: | BOMPR4A |
| NCBI Protein Information: | bone morphogenetic protein 4; BMP-4; BMP-2B; bone morphogenetic protein 2B |
| UniProt Protein Name: | Bone morphogenetic protein 4 |
| UniProt Synonym Protein Names: | Bone morphogenetic protein 2B |
| Protein Family: | Bone morphogenetic protein |
| UniProt Gene Name: | Bmp4 |
| UniProt Entry Name: | BMP4_RAT |
As the RLU values of the standard curve may vary according to the conditions of the actual assay performance (e. g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
| Concentration (pg/mL) | RLU | Average | Corrected |
| 500 | 49989 53085 | 51537 | 51508 |
| 250 | 19386 22610 | 20998 | 20969 |
| 125 | 9774 9018 | 9396 | 9367 |
| 62.5 | 4194 4830 | 4512 | 4483 |
| 31.25 | 2326 2272 | 2299 | 2270 |
| 15.63 | 1287 1213 | 1250 | 1221 |
| 7.81 | 693 787 | 740 | 711 |
| 0 | 28 30 | 29 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Rat BMP-4 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat BMP-4 were tested on 3 different plates, 20 replicates in each plate.
| Intra-assay Precision | Inter-assay Precision | |||||
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 27.06 | 68.94 | 230.38 | 28.78 | 64.06 | 220.64 |
| Standard deviation | 2.22 | 6.11 | 24.37 | 3.00 | 4.68 | 17.94 |
| C V (%) | 8.20 | 8.86 | 10.58 | 10.42 | 7.31 | 8.13 |
Recovery
The recovery of Rat BMP-4 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
| Sample Type | Range (%) | Average Recovery (%) |
| Serum (n=5) | 98-114 | 104 |
| EDTA plasma (n=5) | 89-103 | 95 |
| Cell culture media (n=5) | 87-101 | 94 |
Linearity
Samples were spiked with high concentrations of Rat BMP-4 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
| 1:2 | Range (%) | 97-110 | 88-103 | 93-108 |
| Average (%) | 104 | 95 | 101 | |
| 1:4 | Range (%) | 97-109 | 92-104 | 85-96 |
| Average (%) | 103 | 99 | 91 | |
| 1:8 | Range (%) | 99-111 | 88-102 | 98-113 |
| Average (%) | 105 | 96 | 106 | |
| 1:16 | Range (%) | 89-104 | 99-114 | 88-101 |
| Average (%) | 95 | 106 | 95 |
An unopened kit can be stored at 4°C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.
| Item | Specifications | Storage |
| Micro CLIA Plate(Dismountable) | 8 wells ×12 strips | -20°C, 6 months |
| Reference Standard | 2 vials | |
| Concentrated Biotinylated Detection Ab (100×) | 1 vial, 120 µL | |
| Concentrated HRP Conjugate (100×) | 1 vial, 120 µL | -20°C(shading light), 6 months |
| Reference Standard & Sample Diluent | 1 vial, 20 mL | 4°C, 6 months |
| Biotinylated Detection Ab Diluent | 1 vial, 14 mL | |
| HRP Conjugate Diluent | 1 vial, 14 mL | |
| Concentrated Wash Buffer (25×) | 1 vial, 30 mL | |
| Substrate Reagent A | 1 vial, 5 mL | 4°C (shading light) |
| Substrate Reagent B | 1 vial, 5 mL | 4°C (shading light) |
| Plate Sealer | 5 pieces | |
| Product Description | 1 copy | |
| Certificate of Analysis | 1 copy |
- Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
- Aliquot 100µl of standard solutions into the standard wells.
- Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
- Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids. ) into test sample wells.
- Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
- Aspirate the liquid from each well, do not wash. Immediately add 100µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix. Incubate for 1 hour at 37°C.
- Aspirate or decant the solution from the plate and add 350µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
- Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
- Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
- Add 100µL of Substrate mixture solution to each well. Cover with a new plate seal andincubate for no more than 5 min at 37°C. Protect the plate from light.
- Determine the RLU value of each well immediately.
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