Human ACTN2 (Actinin Alpha 2) CLIA Kit (HUES00191)
- Product Type:
- ELISA Kit
- ELISA Type:
- CLIA Kit
- 96 Assays
- ELISA Type:
- Sample Type:
- Serum, plasma and other biological fluids
- Research Area:
- Cell Biology
|Detection range:||78.13-5000 pg/mL|
|Sample type:||Tissue homogenates|
|Repeatability:||CV < 15%|
|Specificity:||This kit recognizes Human ACTN2 in samples. No significant cross-reactivity or interference between Human ACTN2 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 ACTN2. 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 ACTN2 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 ACTN2, 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 ACTN2. The concentration of Human ACTN2 in the samples can be calculated by comparing the RLU of the samples to the standard curve.
|UniProt Protein Function:||ACTN2: F-actin cross-linking protein which is thought to anchor actin to a variety of intracellular structures. This is a bundling protein. Defects in ACTN2 are the cause of cardiomyopathy dilated type 1AA (CMD1AA). Dilated cardiomyopathy is a disorder characterized by ventricular dilation and impaired systolic function, resulting in congestive heart failure and arrhythmia. Patients are at risk of premature death. Belongs to the alpha-actinin family.|
|UniProt Protein Details:|
Protein type:Motility/polarity/chemotaxis; Cytoskeletal
Chromosomal Location of Human Ortholog: 1q42-q43
Cellular Component: cortical actin cytoskeleton; cytoskeleton; focal adhesion; extracellular region; dendritic spine; actin filament; pseudopodium; Z disc; cytosol; filopodium
Molecular Function:actin filament binding; protein dimerization activity; integrin binding; identical protein binding; protein binding; LIM domain binding; structural constituent of muscle; cytoskeletal protein binding; titin binding; calcium ion binding; FATZ binding; thyroid hormone receptor coactivator activity
Biological Process: focal adhesion formation; platelet activation; negative regulation of potassium ion transport; positive regulation of potassium ion transport; muscle filament sliding; protein homotetramerization; regulation of apoptosis; synaptic transmission; regulation of membrane potential; platelet degranulation; microspike biogenesis; blood coagulation; cell adhesion
Disease: Cardiomyopathy, Dilated, 1aa
|NCBI Summary:||Alpha actinins belong to the spectrin gene superfamily which represents a diverse group of cytoskeletal proteins, including the alpha and beta spectrins and dystrophins. Alpha actinin is an actin-binding protein with multiple roles in different cell types. In nonmuscle cells, the cytoskeletal isoform is found along microfilament bundles and adherens-type junctions, where it is involved in binding actin to the membrane. In contrast, skeletal, cardiac, and smooth muscle isoforms are localized to the Z-disc and analogous dense bodies, where they help anchor the myofibrillar actin filaments. This gene encodes a muscle-specific, alpha actinin isoform that is expressed in both skeletal and cardiac muscles. Several transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, May 2013]|
|NCBI GenInfo Identifier:||543742|
|NCBI Gene ID:||88|
|NCBI Accession:||P35609. 1|
|UniProt Secondary Accession:||P35609,Q86TF4, Q86TI8, B1ANE4, B2RCS5,|
|UniProt Related Accession:||P35609|
|Molecular Weight:||103,920 Da|
|NCBI Full Name:||Alpha-actinin-2|
|NCBI Synonym Full Names:||actinin, alpha 2|
|NCBI Official Symbol:||ACTN2|
|NCBI Official Synonym Symbols:||CMD1AA|
|NCBI Protein Information:||alpha-actinin-2; F-actin cross-linking protein; alpha-actinin skeletal muscle|
|UniProt Protein Name:||Alpha-actinin-2|
|UniProt Synonym Protein Names:||Alpha-actinin skeletal muscle isoform 2; F-actin cross-linking protein|
|UniProt Gene Name:||ACTN2|
|UniProt Entry Name:||ACTN2_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 ACTN2 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human ACTN2 were tested on 3 different plates, 20 replicates in each plate.
|Intra-assay Precision||Inter-assay Precision|
|C V (%)||12.57||8.42||8.34||8.73||7.71||11.07|
The recovery of Human ACTN2 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)||89-102||95|
|Cell culture media (n=5)||84-96||90|
Samples were spiked with high concentrations of Human ACTN2 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.