Human Cell Biology ELISA Kits 1
Human FLNC (Filamin C, Gamma) ELISA Kit (HUES03006)
- SKU:
- HUES03006
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- Q14315
- Sensitivity:
- 0.19ng/mL
- Range:
- 0.31-20ng/mL
- ELISA Type:
- Sandwich
- 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: | Colormetric |
| Detection Range: | 0.31-20 ng/mL |
| Sensitivity: | 0.19 ng/mL |
| Sample Volume Required Per Well: | 100µL |
| Sample Type: | Serum, plasma and other biological fluids |
| Specificity: | This kit recognizes Human FLNC in samples. No significant cross-reactivity or interference between Human FLNC 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 FLNC. 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 FLNC 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 FLNC, 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 FLNC. The concentration of Human FLNC in samples can be calculated by comparing the OD of the samples to the standard curve.
| UniProt Protein Function: | FLNC: Muscle-specific filamin, which plays a central role in muscle cells, probably by functioning as a large actin-cross- linking protein. May be involved in reorganizing the actin cytoskeleton in response to signaling events, and may also display structural functions at the Z lines in muscle cells. Critical for normal myogenesis and for maintaining the structural integrity of the muscle fibers. Defects in FLNC are the cause of myopathy myofibrillar type 5 (MFM5). A neuromuscular disorder, usually with an adult onset, characterized by focal myofibrillar destruction and pathological cytoplasmic protein aggregations, and clinical features of a limb-girdle myopathy. Defects in FLNC are the cause of myopathy distal type 4 (MPD4). MPD4 is a slowly progressive muscular disorder characterized by distal muscle weakness and atrophy affecting the upper and lower limbs. Onset occurs around the third to fourth decades of life, and patients remain ambulatory even after long disease duration. Muscle biopsy shows non-specific changes with no evidence of rods, necrosis, or inflammation. Belongs to the filamin family. 2 isoforms of the human protein are produced by alternative splicing. |
| UniProt Protein Details: | Protein type:Motility/polarity/chemotaxis; Cytoskeletal Chromosomal Location of Human Ortholog: 7q32-q35 Cellular Component: costamere; focal adhesion; cytoskeleton; cytoplasm; plasma membrane; sarcoplasm; Z disc; cytosol; sarcolemma Molecular Function:actin filament binding; protein binding; cytoskeletal protein binding; ankyrin binding Biological Process: muscle fiber development Disease: Myopathy, Myofibrillar, 5; Myopathy, Distal, 4 |
| NCBI Summary: | This gene encodes one of three related filamin genes, specifically gamma filamin. These filamin proteins crosslink actin filaments into orthogonal networks in cortical cytoplasm and participate in the anchoring of membrane proteins for the actin cytoskeleton. Three functional domains exist in filamin: an N-terminal filamentous actin-binding domain, a C-terminal self-association domain, and a membrane glycoprotein-binding domain. Two transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Jul 2008] |
| UniProt Code: | Q14315 |
| NCBI GenInfo Identifier: | 254763419 |
| NCBI Gene ID: | 2318 |
| NCBI Accession: | Q14315. 3 |
| UniProt Secondary Accession: | Q14315,O95303, Q07985, Q9NS12, Q9NYE5, Q9UMR8, Q9Y503 B2ZZ88, |
| UniProt Related Accession: | Q14315 |
| Molecular Weight: | 291,022 Da |
| NCBI Full Name: | Filamin-C |
| NCBI Synonym Full Names: | filamin C, gamma |
| NCBI Official Symbol: | FLNC |
| NCBI Official Synonym Symbols: | ABPA; ABPL; FLN2; MFM5; MPD4; ABP-280; ABP280A |
| NCBI Protein Information: | filamin-C; filamin-2; ABP-280-like protein; ABP-L, gamma filamin; actin binding protein 280 |
| UniProt Protein Name: | Filamin-C |
| UniProt Synonym Protein Names: | ABP-280-like protein; ABP-L; Actin-binding-like protein; Filamin-2; Gamma-filamin |
| Protein Family: | Filamin |
| UniProt Gene Name: | FLNC |
| UniProt Entry Name: | FLNC_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 |
| 20 | 2.371 2.417 | 2.394 | 2.342 |
| 10 | 1.616 1.638 | 1.627 | 1.575 |
| 5 | 0.961 0.921 | 0.941 | 0.889 |
| 2.5 | 0.438 0.46 | 0.449 | 0.397 |
| 1.25 | 0.211 0.205 | 0.208 | 0.156 |
| 0.63 | 0.164 0.136 | 0.15 | 0.098 |
| 0.31 | 0.096 0.108 | 0.102 | 0.05 |
| 0 | 0.049 0.055 | 0.052 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human FLNC were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human FLNC 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) | 1.08 | 3.21 | 9.35 | 1.17 | 3.28 | 9.18 |
| Standard deviation | 0.06 | 0.16 | 0.46 | 0.06 | 0.19 | 0.45 |
| C V (%) | 5.56 | 4.98 | 4.92 | 5.13 | 5.79 | 4.90 |
Recovery
The recovery of Human FLNC 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) | 89-104 | 95 |
| EDTA plasma (n=5) | 85-99 | 90 |
| Cell culture media (n=5) | 91-104 | 99 |
Linearity
Samples were spiked with high concentrations of Human FLNC 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 (%) | 90-103 | 95-111 | 99-111 |
| Average (%) | 95 | 103 | 104 | |
| 1:4 | Range (%) | 90-104 | 87-98 | 84-94 |
| Average (%) | 96 | 93 | 89 | |
| 1:8 | Range (%) | 86-98 | 83-94 | 84-94 |
| Average (%) | 93 | 88 | 89 | |
| 1:16 | Range (%) | 89-104 | 83-97 | 85-99 |
| Average (%) | 95 | 90 | 91 |
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.
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