Human GREM1 (Gremlin 1) CLIA Kit (HUES00786)
- SKU:
- HUES00786
- Product Type:
- ELISA Kit
- ELISA Type:
- CLIA Kit
- Size:
- 96 Assays
- Sensitivity:
- 18.75pg/mL
- Range:
- 31.25-2000pg/mL
- ELISA Type:
- Sandwich
- Synonyms:
- CKTSF1B1, Gremlin 1,Cysteine Knot Superfamily,Homolog(Xenopus Laevis)
- Reactivity:
- Human
- Sample Type:
- Serum, plasma and other biological fluids
- Research Area:
- Cell Biology
Description
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Human |
| Detection method: | Chemiluminescence |
| Detection range: | 31.25-2000 pg/mL |
| Sensitivity: | 18.75 pg/mL |
| Sample volume: | 100µL |
| Sample type: | Serum, plasma and other biological fluids |
| Repeatability: | CV < 15% |
| Specificity: | This kit recognizes Human GREM1 in samples. No significant cross-reactivity or interference between Human GREM1 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 Human GREM1. 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 Human 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 Human GREM1, 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 Human GREM1. The concentration of Human GREM1 in the samples can be calculated by comparing the RLU 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 Chromosomal Location of Human Ortholog: 15q13. 3 Molecular Function:morphogen activity; protein binding; receptor agonist activity; transmembrane receptor protein tyrosine kinase activator activity; vascular endothelial growth factor receptor 2 binding Biological Process: cell migration during sprouting angiogenesis; cell morphogenesis; collagen fibril organization; determination of dorsal identity; 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 chondrocyte differentiation; negative regulation of osteoblast proliferation; positive regulation of cell proliferation; positive regulation of receptor internalization; positive regulation of telomerase activity; positive regulation of transcription from RNA polymerase II promoter; regulation of stress-activated MAPK cascade; signal transduction |
| NCBI Summary: | This gene encodes a member of the BMP (bone morphogenic protein) antagonist family. Like BMPs, BMP antagonists contain cystine knots and typically form homo- and heterodimers. The CAN (cerberus and dan) subfamily of BMP antagonists, to which this gene belongs, is characterized by a C-terminal cystine knot with an eight-membered ring. The antagonistic effect of the secreted glycosylated protein encoded by this gene is likely due to its direct binding to BMP proteins. As an antagonist of BMP, this gene may play a role in regulating organogenesis, body patterning, and tissue differentiation. In mouse, this protein has been shown to relay the sonic hedgehog (SHH) signal from the polarizing region to the apical ectodermal ridge during limb bud outgrowth. Alternatively spliced transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Jul 2010] |
| UniProt Code: | O60565 |
| NCBI GenInfo Identifier: | 62510668 |
| NCBI Gene ID: | 26585 |
| NCBI Accession: | O60565. 1 |
| UniProt Secondary Accession: | O60565,Q52LV3, Q8N914, Q8N936, |
| UniProt Related Accession: | O60565 |
| 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: | DRM; HMPS; MPSH; PIG2; CRAC1; CRCS4; DAND2; HMPS1; IHG-2; DUP15q; C15DUPq; GREMLIN; CKTSF1B1 |
| NCBI Protein Information: | gremlin-1 |
| UniProt Protein Name: | Gremlin-1 |
| UniProt Synonym Protein Names: | Cell proliferation-inducing gene 2 protein; Cysteine knot superfamily 1, BMP antagonist 1; DAN domain family member 2; Down-regulated in Mos-transformed cells protein; Increased in high glucose protein 2; IHG-2 |
| Protein Family: | Gremlin |
| UniProt Gene Name: | GREM1 |
| UniProt Entry Name: | GREM1_HUMAN |
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 |
| 2000 | 52981 58461 | 55721 | 55693 |
| 1000 | 22946 25002 | 23974 | 23946 |
| 500 | 11467 10597 | 11032 | 11004 |
| 250 | 5246 5342 | 5294 | 5266 |
| 125 | 2757 2459 | 2608 | 2580 |
| 62.5 | 1405 1217 | 1311 | 1283 |
| 31.25 | 667 681 | 674 | 646 |
| 0 | 28 28 | 28 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human GREM1 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human 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) | 95.99 | 259.38 | 913.08 | 89.99 | 235.01 | 883.20 |
| Standard deviation | 10.10 | 23.50 | 82.45 | 7.33 | 26.23 | 86.29 |
| C V (%) | 10.52 | 9.06 | 9.03 | 8.15 | 11.16 | 9.77 |
Recovery
The recovery of Human 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) | 98-112 | 104 |
| EDTA plasma (n=5) | 89-99 | 94 |
| Cell culture media (n=5) | 100-119 | 108 |
Linearity
Samples were spiked with high concentrations of Human 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 (%) | 94-109 | 93-109 | 87-98 |
| Average (%) | 100 | 101 | 93 | |
| 1:4 | Range (%) | 92-109 | 90-104 | 97-108 |
| Average (%) | 100 | 97 | 102 | |
| 1:8 | Range (%) | 97-112 | 101-115 | 89-103 |
| Average (%) | 104 | 108 | 96 | |
| 1:16 | Range (%) | 96-112 | 101-117 | 101-113 |
| Average (%) | 102 | 109 | 107 |
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.
Related Products
Rat GREM1 (GremLin 1) CLIA Kit (RTES00263)
