Description
Mouse RELN (Reelin) ELISA kit
The Mouse RELN (Reelin) ELISA Kit is a reliable and accurate tool designed for the detection of reelin levels in mouse serum, plasma, and cell culture supernatants. With high sensitivity and specificity, this kit ensures consistent and reproducible results, making it ideal for various research applications.Reelin is a critical protein that plays a key role in regulating neuronal migration and positioning in the brain. Dysregulation of reelin has been implicated in various neurological disorders such as schizophrenia, autism, and Alzheimer's disease.
Therefore, this ELISA kit is essential for studying the role of reelin in these conditions and developing potential therapeutic interventions.Overall, the Mouse RELN (Reelin) ELISA Kit provides researchers with a valuable tool for accurately measuring reelin levels, advancing our understanding of neurological disorders, and potentially identifying new treatment strategies.
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Mouse |
| Detection Method: | Colormetric |
| Detection Range: | 15.63-1000 pg/mL |
| Sensitivity: | 9.38 pg/mL |
| Sample Volume Required Per Well: | 100µL |
| Sample Type: | Serum, plasma and other biological fluids |
| Specificity: | This kit recognizes Mouse RELN in samples. No significant cross-reactivity or interference between Mouse RELN 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 RELN. 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 RELN 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 RELN, 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 RELN. The concentration of Mouse RELN in samples can be calculated by comparing the OD of the samples to the standard curve.
| UniProt Protein Function: | reelin: Extracellular matrix serine protease that plays a role in layering of neurons in the cerebral cortex and cerebellum. Regulates microtubule function in neurons and neuronal migration. Affects migration of sympathetic preganglionic neurons in the spinal cord, where it seems to act as a barrier to neuronal migration. Enzymatic activity is important for the modulation of cell adhesion. Binding to the extracellular domains of lipoprotein receptors VLDLR and LRP8/APOER2 induces tyrosine phosphorylation of DAB1 and modulation of TAU phosphorylation. Defects in RELN are the cause of lissencephaly type 2 (LIS2); also known as lissencephaly with cerebellar hypoplasia or Norman-Roberts syndrome. LIS2 is a classic type lissencephaly associated with abnormalities of the cerebellum, hippocampus and brainstem. Individuals with LIS2 are severely ataxic, mentally retarded and suffer from epilepsy. Belongs to the reelin family. 3 isoforms of the human protein are produced by alternative splicing. |
| UniProt Protein Details: | Protein type:Motility/polarity/chemotaxis; Cell development/differentiation; Cell adhesion; EC 3. 4. 21. -; Secreted; Secreted, signal peptide; Protease Cellular Component: extracellular space; proteinaceous extracellular matrix; neuron projection; cell soma; axon; dendrite; cytoplasm; extracellular region; perikaryon Molecular Function:peptidase activity; protein binding; hydrolase activity; serine-type peptidase activity; metal ion binding Biological Process: axon guidance; cellular morphogenesis during differentiation; peptidyl-tyrosine phosphorylation; central nervous system development; spinal cord patterning; multicellular organismal development; neuron migration; response to pain; locomotory behavior; proteolysis; positive regulation of synaptic transmission, glutamatergic; glial cell differentiation; forebrain development; dendrite development; cell adhesion; layer formation in the cerebral cortex; associative learning; positive regulation of TOR signaling pathway; thyroid hormone metabolic process; regulation of behavior; regulation of synaptic transmission; cell migration; dentate gyrus development; cerebral cortex tangential migration; hippocampus development; activation of CREB transcription factor; positive regulation of phosphoinositide 3-kinase cascade; positive regulation of small GTPase mediated signal transduction; positive regulation of peptidyl-tyrosine phosphorylation; long-term memory; regulation of gene expression; positive regulation of protein kinase activity; cerebral cortex development; brain development |
| UniProt Code: | Q60841 |
| NCBI GenInfo Identifier: | 117320554 |
| NCBI Gene ID: | 19699 |
| NCBI Accession: | NP_035391. 2 |
| UniProt Secondary Accession: | Q60841,Q9CUA6, E9PZ78, |
| UniProt Related Accession: | Q60841 |
| Molecular Weight: | 41. 8kDa |
| NCBI Full Name: | reelin |
| NCBI Synonym Full Names: | reelin |
| NCBI Official Symbol: | Reln |
| NCBI Official Synonym Symbols: | rl; reeler |
| NCBI Protein Information: | reelin; extracellular matrix serine protease |
| UniProt Protein Name: | Reelin |
| UniProt Synonym Protein Names: | Reeler protein |
| UniProt Gene Name: | Reln |
| UniProt Entry Name: | RELN_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 |
| 1000 | 2.352 2.366 | 2.359 | 2.279 |
| 500 | 1.686 1.702 | 1.694 | 1.614 |
| 250 | 0.971 0.951 | 0.961 | 0.881 |
| 125 | 0.504 0.516 | 0.51 | 0.43 |
| 62.5 | 0.297 0.285 | 0.291 | 0.211 |
| 31.25 | 0.188 0.174 | 0.181 | 0.101 |
| 15.63 | 0.132 0.132 | 0.132 | 0.052 |
| 0 | 0.075 0.085 | 0.08 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse RELN were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse RELN 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) | 50.59 | 107.00 | 492.54 | 55.00 | 104.73 | 541.46 |
| Standard deviation | 3.41 | 5.21 | 15.12 | 3.32 | 4.69 | 25.02 |
| C V (%) | 6.74 | 4.87 | 3.07 | 6.04 | 4.48 | 4.62 |
Recovery
The recovery of Mouse RELN 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) | 84-97 | 91 |
| EDTA plasma (n=5) | 94-106 | 99 |
| Cell culture media (n=5) | 87-104 | 95 |
Linearity
Samples were spiked with high concentrations of Mouse RELN 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 (%) | 93-108 | 89-99 | 97-110 |
| Average (%) | 101 | 94 | 104 | |
| 1:4 | Range (%) | 91-103 | 80-91 | 82-95 |
| Average (%) | 96 | 86 | 88 | |
| 1:8 | Range (%) | 93-104 | 87-100 | 84-96 |
| Average (%) | 98 | 92 | 91 | |
| 1:16 | Range (%) | 85-98 | 82-91 | 88-100 |
| Average (%) | 92 | 86 | 93 |
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.
