Human ACT alpha1 (Actin Alpha 1, Skeletal Muscle) CLIA Kit (HUES00194)
- Product Type:
- CLIA Kit
- 96 Assays
- ELISA Type:
- ACTA1, ASMA, CFTD, CFTD1, CFTDM, MPFD, NEM1, NEM2, NEM3
- Tested Sample Types:
- Serum, plasma and other biological fluids
|Detection range:||62.50-4000 pg/mL|
|Sample type:||Serum, plasma and other biological fluids|
|Repeatability:||CV < 15%|
|Specificity:||This kit recognizes Human ACT alpha1 in samples. No significant cross-reactivity or interference between Human ACT alpha1 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 ACT alpha1. 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 ACT alpha1 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 ACT alpha1, 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 ACT alpha1. The concentration of Human ACT alpha1 in the samples can be calculated by comparing the RLU of the samples to the standard curve.
|UniProt Protein Function:||ACTA1: Actins are highly conserved proteins that are involved in various types of cell motility and are ubiquitously expressed in all eukaryotic cells. Polymerization of globular actin (G-actin) leads to a structural filament (F-actin) in the form of a two-stranded helix. Each actin can bind to 4 others. Interacts with TTID. Interacts (via its C-terminus) with USP25; the interaction occurs for all USP25 isoforms but is strongest for isoform USP25m in muscle differentiating cells. Belongs to the actin family.|
|UniProt Protein Details:|
Protein type:Cell development/differentiation; Motility/polarity/chemotaxis; Contractile; Cytoskeletal
Chromosomal Location of Human Ortholog: 1q42. 13
Cellular Component: extracellular space; sarcomere; stress fiber; actin filament; cytosol; striated muscle thin filament; actin cytoskeleton
Molecular Function:protein binding; myosin binding; structural constituent of cytoskeleton; ADP binding; ATP binding
Biological Process: skeletal muscle fiber adaptation; muscle contraction; muscle thin filament assembly; response to steroid hormone stimulus; response to mechanical stimulus; response to lithium ion; response to extracellular stimulus; skeletal muscle fiber development; cell growth; muscle filament sliding
Disease: Nemaline Myopathy 3; Myopathy, Congenital, With Fiber-type Disproportion
|NCBI Summary:||The product encoded by this gene belongs to the actin family of proteins, which are highly conserved proteins that play a role in cell motility, structure and integrity. Alpha, beta and gamma actin isoforms have been identified, with alpha actins being a major constituent of the contractile apparatus, while beta and gamma actins are involved in the regulation of cell motility. This actin is an alpha actin that is found in skeletal muscle. Mutations in this gene cause nemaline myopathy type 3, congenital myopathy with excess of thin myofilaments, congenital myopathy with cores, and congenital myopathy with fiber-type disproportion, diseases that lead to muscle fiber defects. [provided by RefSeq, Jul 2008]|
|NCBI GenInfo Identifier:||4501881|
|NCBI Gene ID:||58|
|NCBI Accession:||NP_001091. 1|
|UniProt Secondary Accession:||P68133,P02568, P99020, Q5T8M9,|
|UniProt Related Accession:||P68133|
|Molecular Weight:||42,051 Da|
|NCBI Full Name:||actin, alpha skeletal muscle|
|NCBI Synonym Full Names:||actin, alpha 1, skeletal muscle|
|NCBI Official Symbol:||ACTA1|
|NCBI Official Synonym Symbols:||ACTA; ASMA; CFTD; MPFD; NEM1; NEM2; NEM3; CFTD1; CFTDM|
|NCBI Protein Information:||actin, alpha skeletal muscle; nemaline myopathy type 3|
|UniProt Protein Name:||Actin, alpha skeletal muscle|
|UniProt Synonym Protein Names:||Alpha-actin-1|
|UniProt Gene Name:||ACTA1|
|UniProt Entry Name:||ACTS_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.
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human ACT alpha1 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human ACT alpha1 were tested on 3 different plates, 20 replicates in each plate.
|Intra-assay Precision||Inter-assay Precision|
|C V (%)||11.10||10.65||6.32||12.80||9.98||9.66|
The recovery of Human ACT alpha1 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
|Sample Type||Range (%)||Average Recovery (%)|
|EDTA plasma (n=5)||96-112||103|
|Cell culture media (n=5)||86-98||92|
Samples were spiked with high concentrations of Human ACT alpha1 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)|
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.
|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.