Human CTNN alpha1 (Catenin, Alpha 1) ELISA Kit (HUES01823)
- Product Type:
- ELISA Kit
- 96 Assays
- ELISA Type:
- Tested Sample Types:
- Serum, plasma and other biological fluids
|Detection Range:||0.31-20 ng/mL|
|Sample Volume Required Per Well:||100µL|
|Sample Type:||Serum, plasma and other biological fluids|
|Specificity:||This kit recognizes Human CTNN alpha1 in samples. No significant cross-reactivity or interference between Human CTNN alpha1 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 CTNN alpha1. 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 CTNN 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 CTNN alpha1, 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 CTNN alpha1. The concentration of Human CTNN alpha1 in samples can be calculated by comparing the OD of the samples to the standard curve.
|UniProt Protein Function:||CTNNA1: Associates with the cytoplasmic domain of a variety of cadherins. The association of catenins to cadherins produces a complex which is linked to the actin filament network, and which seems to be of primary importance for cadherins cell-adhesion properties. Can associate with both E- and N-cadherins. Originally believed to be a stable component of E-cadherin/catenin adhesion complexes and to mediate the linkage of cadherins to the actin cytoskeleton at adherens junctions. In contrast, cortical actin was found to be much more dynamic than E-cadherin/catenin complexes and CTNNA1 was shown not to bind to F-actin when assembled in the complex suggesting a different linkage between actin and adherens junctions components. The homodimeric form may regulate actin filament assembly and inhibit actin branching by competing with the Arp2/3 complex for binding to actin filaments. May play a crucial role in cell differentiation. Monomer and homodimer; the monomer preferentially binds to CTNNB1 and the homodimer to actin. Binds MLLT4 and F-actin. Possible component of an E-cadherin/ catenin adhesion complex together with E-cadherin/CDH1 and beta-catenin/CTNNB1 or gamma- catenin/JUP; the complex is located to adherens junctions. The stable association of CTNNA1 is controversial as CTNNA1 was shown not to bind to F-actin when assembled in the complex. Alternatively, the CTNNA1-containing complex may be linked to F- actin by other proteins such as LIMA1. Interacts with ARHGAP21 and with AJUBA. Interacts with LIMA1. Expressed ubiquitously in normal tissues. Belongs to the vinculin/alpha-catenin family. 2 isoforms of the human protein are produced by alternative splicing.|
|UniProt Protein Details:|
Protein type:Motility/polarity/chemotaxis; Actin-binding
Chromosomal Location of Human Ortholog: 5q31. 2
Cellular Component: focal adhesion; lamellipodium; acrosome; plasma membrane; catenin complex; zonula adherens; intercellular junction; cytosol; actin cytoskeleton
Molecular Function:actin filament binding; protein binding; cadherin binding; gamma-catenin binding; beta-catenin binding; structural molecule activity; vinculin binding
Biological Process: intercellular junction assembly and maintenance; apical junction assembly; gap junction assembly; protein heterooligomerization; axon regeneration; male gonad development; establishment and/or maintenance of cell polarity; actin filament organization; odontogenesis of dentine-containing teeth; muscle cell differentiation; negative regulation of neuroblast proliferation; ovarian follicle development; response to estrogen stimulus; positive regulation of smoothened signaling pathway; positive regulation of muscle cell differentiation; cell adhesion; vascular endothelial growth factor receptor signaling pathway; aging
|NCBI GenInfo Identifier:||461853|
|NCBI Gene ID:||1495|
|NCBI Accession:||P35221. 1|
|UniProt Secondary Accession:||P35221,Q12795, Q8N1C0,|
|UniProt Related Accession:||P35221|
|Molecular Weight:||59,550 Da|
|NCBI Full Name:||Catenin alpha-1|
|NCBI Synonym Full Names:||catenin (cadherin-associated protein), alpha 1, 102kDa|
|NCBI Official Symbol:||CTNNA1|
|NCBI Official Synonym Symbols:||CAP102|
|NCBI Protein Information:||catenin alpha-1; alpha-catenin; alphaE-catenin; alpha E-catenin; alpha-E-catenin; renal carcinoma antigen NY-REN-13; cadherin-associated protein,102kDa|
|UniProt Protein Name:||Catenin alpha-1|
|UniProt Synonym Protein Names:||Alpha E-catenin; Cadherin-associated protein; Renal carcinoma antigen NY-REN-13|
|UniProt Gene Name:||CTNNA1|
|UniProt Entry Name:||CTNA1_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.
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human CTNN 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 CTNN alpha1 were tested on 3 different plates, 20 replicates in each plate.
|Intra-assay Precision||Inter-assay Precision|
|C V (%)||6.67||3.96||4.77||5.45||5.43||3.30|
The recovery of Human CTNN 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)||85-96||91|
|Cell culture media (n=5)||86-98||93|
Samples were spiked with high concentrations of Human CTNN 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 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 100 µL of standard solutions into the standard wells.
- Add 100 µL of Sample / Standard dilution buffer into the control (zero) well.
- Add 100 µL 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 100 µL 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 350 µL 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 100 µL 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 90 µL 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.