Description
Mouse GREM1 (GremLin 1) ELISA Kit
The Mouse GREM1 (Gremlin 1) ELISA Kit is specifically designed for the precise measurement of GREM1 levels in mouse serum, plasma, and cell culture supernatants. This kit offers exceptional sensitivity and specificity, guaranteeing accurate and consistent results, making it perfect for various research applications.GREM1, also known as Gremlin 1, is a key regulator in the BMP (Bone Morphogenetic Protein) signaling pathway, playing a vital role in embryonic development, tissue regeneration, and disease progression.
With its involvement in various biological processes, GREM1 serves as a valuable biomarker for studying developmental disorders, cancer, and fibrosis, providing insights for potential therapeutic strategies.Overall, the Mouse GREM1 ELISA Kit is a reliable tool for researchers looking to investigate the role of GREM1 in disease pathogenesis and therapeutic interventions in mouse models.
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Mouse |
| Detection Method: | Colormetric |
| Detection Range: | 78.13-5000 pg/mL |
| Sensitivity: | 46.88 pg/mL |
| Sample Volume Required Per Well: | 100µL |
| Sample Type: | Serum, plasma and other biological fluids |
| Specificity: | This kit recognizes Mouse GREM1 in samples. No significant cross-reactivity or interference between Mouse GREM1 and analogues was observed. |
This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Mouse GREM1. Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Mouse GREM1 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 Mouse GREM1, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Mouse GREM1. The concentration of Mouse GREM1 in samples can be calculated by comparing the OD of the samples to the standard curve.
| UniProt Protein Function: | GREM1: Cytokine that may play an important role during carcinogenesis and metanephric kidney organogenesis, as a BMP antagonist required for early limb outgrowth and patterning in maintaining the FGF4-SHH feedback loop. Down-regulates the BMP4 signaling in a dose-dependent manner. Acts as inhibitor of monocyte chemotaxis. Interacts with SLIT1 and SLIT2 in a glycosylation- dependent manner. By high glucose through TGFB1-mediated pathways in mesangial cell. Down-regulated in tumor cell lines. Highly expressed in small intestine, fetal brain and colon. Weakly expressed in brain, ovary, prostate, pancreas and skeletal muscle. In brain found in the region localized around the internal capsule in the large subcortical nuclei, including caudate, putamen, substantia nigra, thalamus and subthalamus. Predominantly expressed in normal cells including neurons, astrocytes and fibroblasts. Belongs to the DAN family. 2 isoforms of the human protein are produced by alternative splicing. |
| UniProt Protein Details: | Protein type:Secreted, signal peptide; Secreted Cellular Component: cell surface; extracellular region; extracellular space Molecular Function:cytokine activity; heparan sulfate proteoglycan binding; receptor agonist activity; transmembrane receptor protein tyrosine kinase activator activity; vascular endothelial growth factor receptor 2 binding Biological Process: activation of NF-kappaB transcription factor; angiogenesis; cell migration during sprouting angiogenesis; cell morphogenesis; cell-cell signaling; collagen fibril organization; determination of dorsal identity; embryonic limb morphogenesis; endothelial cell migration; limb development; negative regulation of apoptosis; negative regulation of BMP signaling pathway; negative regulation of bone mineralization; negative regulation of bone remodeling; negative regulation of cell growth; negative regulation of chondrocyte differentiation; negative regulation of leukocyte chemotaxis; negative regulation of osteoblast proliferation; negative regulation of transcription, DNA-dependent; organ morphogenesis; positive regulation of angiogenesis; positive regulation of cell proliferation; positive regulation of NF-kappaB import into nucleus; positive regulation of receptor internalization; positive regulation of telomerase activity; positive regulation of transcription from RNA polymerase II promoter; positive regulation of transcription, DNA-dependent; proximal/distal pattern formation; regulation of focal adhesion formation; regulation of stress-activated MAPK cascade; signal transduction; ureteric bud branching |
| UniProt Code: | O70326 |
| NCBI GenInfo Identifier: | 62510669 |
| NCBI Gene ID: | 23892 |
| NCBI Accession: | O70326. 1 |
| UniProt Related Accession: | O70326 |
| Molecular Weight: | |
| NCBI Full Name: | Gremlin-1 |
| NCBI Synonym Full Names: | gremlin 1, DAN family BMP antagonist |
| NCBI Official Symbol: | Grem1 |
| NCBI Official Synonym Symbols: | ld; Drm; Grem; Cktsf1b1 |
| NCBI Protein Information: | gremlin-1 |
| UniProt Protein Name: | Gremlin-1 |
| UniProt Synonym Protein Names: | Cysteine knot superfamily 1, BMP antagonist 1; Down-regulated in Mos-transformed cells protein |
| Protein Family: | Gremlin |
| UniProt Gene Name: | Grem1 |
| UniProt Entry Name: | GREM1_MOUSE |
As the OD 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) | O.D | Average | Corrected |
| 5000 | 2.538 2.572 | 2.555 | 2.48 |
| 2500 | 1.77 1.792 | 1.781 | 1.706 |
| 1250 | 1.031 1.011 | 1.021 | 0.946 |
| 625 | 0.507 0.507 | 0.507 | 0.432 |
| 312.5 | 0.279 0.273 | 0.276 | 0.201 |
| 156.25 | 0.197 0.169 | 0.183 | 0.108 |
| 78.13 | 0.13 0.13 | 0.13 | 0.055 |
| 0 | 0.069 0.081 | 0.075 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse GREM1 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse GREM1 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) | 255.76 | 529.21 | 2079.55 | 275.81 | 516.60 | 2005.58 |
| Standard deviation | 15.96 | 29.69 | 80.69 | 17.60 | 29.39 | 83.43 |
| C V (%) | 6.24 | 5.61 | 3.88 | 6.38 | 5.69 | 4.16 |
Recovery
The recovery of Mouse GREM1 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) | 90-104 | 96 |
| EDTA plasma (n=5) | 96-107 | 101 |
| Cell culture media (n=5) | 86-98 | 92 |
Linearity
Samples were spiked with high concentrations of Mouse GREM1 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 (%) | 96-109 | 93-109 | 97-112 |
| Average (%) | 104 | 100 | 104 | |
| 1:4 | Range (%) | 90-102 | 82-96 | 83-95 |
| Average (%) | 97 | 88 | 90 | |
| 1:8 | Range (%) | 91-104 | 80-91 | 81-96 |
| Average (%) | 97 | 86 | 87 | |
| 1:16 | Range (%) | 92-109 | 80-93 | 86-100 |
| Average (%) | 100 | 86 | 92 |
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 ELISA 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 | 1 vial, 10 mL | 4°C(shading light) |
| Stop Solution | 1 vial, 10 mL | 4°C |
| 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 90µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37°C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
- Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
- Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.
