Description
Human SPTAN1 (Alpha-Fodrin) ELISA Kit
The Human SPTAN1 (Alpha Fodrin) ELISA Kit is a powerful tool for precise quantification of SPTAN1 levels in human samples including serum, plasma, and cell culture supernatants. With its exceptional sensitivity and specificity, researchers can trust the accuracy and consistency of their results, making this kit suitable for a variety of research applications.SPTAN1, also known as Alpha Fodrin, is a key protein involved in cell structure and function.
Its role in cytoskeleton organization and cellular signaling makes it a valuable biomarker for studying diseases such as cancer, neurological disorders, and autoimmune conditions. By measuring SPTAN1 levels, researchers can gain important insights into disease mechanisms and potentially identify new therapeutic targets.
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Human |
| Detection Method: | Colormetric |
| Detection Range: | 0.16-10 ng/mL |
| Sensitivity: | 0.10 ng/mL |
| Sample Volume Required Per Well: | 100µL |
| Sample Type: | Serum, plasma and other biological fluids |
| Specificity: | This kit recognizes Human SPTAN1 in samples. No significant cross-reactivity or interference between Human SPTAN1 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 Human SPTAN1. 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 Human SPTAN1 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 SPTAN1, 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 Human SPTAN1. The concentration of Human SPTAN1 in samples can be calculated by comparing the OD of the samples to the standard curve.
| UniProt Protein Function: | SPTAN1: a cytoskeletal protein which seems to be involved in secretion, interacts with calmodulin in a calcium-dependent manner and is thus candidate for the calcium-dependent movement of the cytoskeleton at the membrane. Capable to forming dimers which can further associate to tetramers. |
| UniProt Protein Details: | Protein type:Cytoskeletal; Motility/polarity/chemotaxis; Adaptor/scaffold Chromosomal Location of Human Ortholog: 9q34. 11 Cellular Component: cytosol; fascia adherens; intracellular membrane-bound organelle; lateral plasma membrane; membrane; microtubule cytoskeleton; protein complex; spectrin; Z disc Molecular Function:actin binding; calcium ion binding; calmodulin binding; protein binding; protein complex binding; protein heterodimerization activity; protein N-terminus binding; structural constituent of cytoskeleton; syntaxin binding Biological Process: actin cytoskeleton reorganization; actin filament capping; activation of MAPKK activity; apoptosis; axon guidance; cell structure disassembly during apoptosis; cellular protein metabolic process; epidermal growth factor receptor signaling pathway; ER to Golgi vesicle-mediated transport; fibroblast growth factor receptor signaling pathway; innate immune response; insulin receptor signaling pathway; MAPKKK cascade; nerve growth factor receptor signaling pathway; post-translational protein modification; programmed cell death; protein amino acid N-linked glycosylation via asparagine; Ras protein signal transduction; small GTPase mediated signal transduction; vascular endothelial growth factor receptor signaling pathway Disease: Epileptic Encephalopathy, Early Infantile, 5 |
| NCBI Summary: | Spectrins are a family of filamentous cytoskeletal proteins that function as essential scaffold proteins that stabilize the plasma membrane and organize intracellular organelles. Spectrins are composed of alpha and beta dimers that associate to form tetramers linked in a head-to-head arrangement. This gene encodes an alpha spectrin that is specifically expressed in nonerythrocytic cells. The encoded protein has been implicated in other cellular functions including DNA repair and cell cycle regulation. Mutations in this gene are the cause of early infantile epileptic encephalopathy-5. Alternate splicing results in multiple transcript variants. [provided by RefSeq, Sep 2010] |
| UniProt Code: | Q13813 |
| NCBI GenInfo Identifier: | 94730425 |
| NCBI Gene ID: | 6709 |
| NCBI Accession: | Q13813. 3 |
| UniProt Secondary Accession: | Q13813,Q13186, Q15324, Q16606, Q59EF1, Q5VXV5, Q5VXV6 Q7Z6M5, Q9P0V0, |
| UniProt Related Accession: | Q13813 |
| Molecular Weight: | 282,282 Da |
| NCBI Full Name: | Spectrin alpha chain, non-erythrocytic 1 |
| NCBI Synonym Full Names: | spectrin alpha, non-erythrocytic 1 |
| NCBI Official Symbol: | SPTAN1 |
| NCBI Official Synonym Symbols: | NEAS; EIEE5; SPTA2 |
| NCBI Protein Information: | spectrin alpha chain, non-erythrocytic 1 |
| UniProt Protein Name: | Spectrin alpha chain, non-erythrocytic 1 |
| UniProt Synonym Protein Names: | Alpha-II spectrin; Fodrin alpha chain; Spectrin, non-erythroid alpha subunit |
| Protein Family: | Spectrin |
| UniProt Gene Name: | SPTAN1 |
| UniProt Entry Name: | SPTN1_HUMAN |
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 (ng/mL) | O.D | Average | Corrected |
| 10 | 2.549 2.565 | 2.557 | 2.499 |
| 5 | 1.643 1.685 | 1.664 | 1.606 |
| 2.5 | 0.993 0.989 | 0.991 | 0.933 |
| 1.25 | 0.443 0.469 | 0.456 | 0.398 |
| 0.63 | 0.27 0.26 | 0.265 | 0.207 |
| 0.32 | 0.173 0.165 | 0.169 | 0.111 |
| 0.16 | 0.11 0.118 | 0.114 | 0.056 |
| 0 | 0.055 0.061 | 0.058 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human SPTAN1 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human SPTAN1 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 (ng/mL) | 0.47 | 1.10 | 3.47 | 0.45 | 1.13 | 3.64 |
| Standard deviation | 0.03 | 0.05 | 0.18 | 0.02 | 0.06 | 0.13 |
| C V (%) | 6.38 | 4.55 | 5.19 | 4.44 | 5.31 | 3.57 |
Recovery
The recovery of Human SPTAN1 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) | 94-110 | 101 |
| EDTA plasma (n=5) | 89-102 | 95 |
| Cell culture media (n=5) | 94-111 | 101 |
Linearity
Samples were spiked with high concentrations of Human SPTAN1 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 (%) | 86-99 | 92-109 | 96-110 |
| Average (%) | 91 | 100 | 102 | |
| 1:4 | Range (%) | 89-101 | 83-97 | 85-100 |
| Average (%) | 94 | 89 | 92 | |
| 1:8 | Range (%) | 89-101 | 85-97 | 88-100 |
| Average (%) | 94 | 92 | 94 | |
| 1:16 | Range (%) | 89-103 | 82-93 | 81-95 |
| Average (%) | 95 | 87 | 88 |
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.
