Human CSNK2A1 (Casein Kinase 2, alpha 1 Polypeptide) ELISA Kit (HUES03467)

SKU:
HUES03467
Product type:
ELISA
Reactivity:
Human
ELISA Type:
Sandwich
$647

Description

ELISA Kit Technical ManualMSDS

Human CSNK2A1 (Casein Kinase 2, alpha 1 Polypeptide) ELISA Kit

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 CSNK2A1 . 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 CSNK2A1 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 CSNK2A1, 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 CSNK2A1. The concentration of Human CSNK2A1 in samples can be calculated by comparing the OD of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityHuman
Detection MethodColormetric
Detection Range0.16-10 ng/mL
Sensitivity0.10 ng/mL
Sample Volume Required Per Well100uL
Sample TypeSerum, plasma and other biological fluids

Specificity

This kit recognizes Human CSNK2A1 in samples. No significant cross-reactivity or interference between Human CSNK2A1 and analogues was observed.

Typical Data

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
102.411
2.417
2.4142.34
51.683
1.697
1.691.616
2.50.985
0.953
0.9690.895
1.250.474
0.494
0.4840.41
0.630.276
0.252
0.2640.19
0.320.186
0.174
0.180.106
0.160.121
0.135
0.1280.054
00.065
0.083
0.074--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human CSNK2A1 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human CSNK2A1 were tested on 3 different plates, 20 replicates in each plate.

Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (ng/mL)0.481.473.400.501.383.15
Standard deviation0.030.080.140.030.070.16
C V (%)6.255.444.126.005.075.08

Recovery

The recovery of Human CSNK2A1 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)88-10294
EDTA plasma (n=5)93-108100
Cell culture media (n=5)89-10296

Linearity

Samples were spiked with high concentrations of Human CSNK2A1 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)
1:2Range (%)89-10292-10395-109
Average (%)9797102
1:4Range (%)87-9985-9784-95
Average (%)929089
1:8Range (%)85-9780-9484-96
Average (%)928691
1:16Range (%)89-10285-9985-99
Average (%)959292

Kit Components & Storage

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 X12 strips-20'C, 6 months
Reference Standard2 vials
Concentrated Biotinylated Detection Ab (100X)1 vial, 120 uL
Concentrated HRP Conjugate (100X)1 vial, 120 uL-20'C(shading light), 6 months
Reference Standard & Sample Diluent1 vial, 20 mL4'C, 6 months
Biotinylated Detection Ab Diluent1 vial, 14 mL
HRP Conjugate Diluent1 vial, 14 mL
Concentrated Wash Buffer (25X)1 vial, 30 mL
Substrate Reagent1 vial, 10 mL4'C(shading light)
Stop Solution1 vial, 10 mL4'C
Plate Sealer5 pieces
Product Description1 copy
Certificate of Analysis1 copy

Human CSNK2A1 (Casein Kinase 2, alpha 1 Polypeptide) ELISA Kit (HUES03467) Assay procedure

    1. 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.
    2. Aliquot 100µl of standard solutions into the standard wells.
    3. Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
    4. Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids) into test sample wells.
    5. Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
    6. 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.
    7. 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.
    8. Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
    9. Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
    10. 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.
    11. Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
    12. Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.

Human CSNK2A1 (Casein Kinase 2, alpha 1 Polypeptide) ELISA Kit (HUES03467) Protein Information

UniProt Protein Function:CK2A1: an ubiquitous protein kinase of the CK2 family. Exists as a tetramer composed of two catalytic subunits, alpha and alpha-prime, and two regulatory beta subunits. The beta subunits undergo autophosphorylation. The isoforms are rarely specified in publications. Two splice variant isoforms have been found. Participates in Wnt signaling. Phosphorylates E2 ubiquitin conjugating enzyme UBC3B inducing its interaction with beta-TRCp and enhancing beta-catenin degradation. May control IkB-alpha and p27Kip1 degradation. component of a CK2-SPT16-SSRP1 complex composed of SSRP1, PT16 CK2-A1, CK2-A2 and CK2-B the complex associating following UV irradiation. Interacts with RNPS1. Mouse transgene causes mammary gland hyperplasia and lymphoma, and activation by bovine parasites leads to fatal lymphoproliferation. Expression and activity are elevated in lung tumors and breast tumors. Antisense drives apoptosis of tumor cell lines and xenografts. Involved in DNA break repair by phosphorylation of scaffold protein XRCC1, phosphorylation of BRCA1, and phosphorylation of p53 in response to UV irradiation. Drosophila CK2 (Timekeeper) is involved in circadian regulation. Phosphorylates and binds to a major component of the inclusion bodies seen in Parkinson?s patients. Inhibitors: 4,5,6,7-tetrabromobenzotriazole (TBB).
UniProt Protein Details:

Protein type:EC 2.7.11.1; Kinase, protein; Protein kinase, Ser/Thr (non-receptor); Protein kinase, Other; Other group; CK2 family

Chromosomal Location of Human Ortholog: 20p13

Cellular Component: Sin3 complex; plasma membrane; PcG protein complex; NuRD complex; cytosol; nucleus

Molecular Function:protein serine/threonine kinase activity; protein binding; protein N-terminus binding; Hsp90 protein binding; ATP binding

Biological Process: axon guidance; Wnt receptor signaling pathway; transcription, DNA-dependent; rhythmic process; negative regulation of caspase activity; signal transduction; positive regulation of cell growth; protein amino acid phosphorylation; regulation of transcription, DNA-dependent; positive regulation of cell proliferation; positive regulation of protein catabolic process; mitotic cell cycle; positive regulation of Wnt receptor signaling pathway

NCBI Summary:Casein kinase II is a serine/threonine protein kinase that phosphorylates acidic proteins such as casein. It is involved in various cellular processes, including cell cycle control, apoptosis, and circadian rhythm. The kinase exists as a tetramer and is composed of an alpha, an alpha-prime, and two beta subunits. The alpha subunits contain the catalytic activity while the beta subunits undergo autophosphorylation. The protein encoded by this gene represents the alpha subunit. While this gene is found on chromosome 20, a related transcribed pseudogene is found on chromosome 11. Three transcript variants encoding two different proteins have been found for this gene. [provided by RefSeq, Jul 2014]
UniProt Code:P68400
NCBI GenInfo Identifier:55977123
NCBI Gene ID:1457
NCBI Accession:P68400.1
UniProt Secondary Accession:P68400,P19138, P20426, Q14013, Q5U065, B4DYS6, D3DVV8
UniProt Related Accession:P68400,Q8NEV1
Molecular Weight:391
NCBI Full Name:Casein kinase II subunit alpha
NCBI Synonym Full Names:casein kinase 2, alpha 1 polypeptide
NCBI Official Symbol:CSNK2A1
NCBI Official Synonym Symbols:CKII; CK2A1; CSNK2A3
NCBI Protein Information:casein kinase II subunit alpha; CK II alpha 3; protein kinase CK2; CK2 catalytic subunit alpha; casein kinase II alpha 1 subunit; casein kinase II alpha 1 polypeptide pseudogene
UniProt Protein Name:Casein kinase II subunit alpha
Protein Family:Casein kinase
UniProt Gene Name:CSNK2A1
UniProt Entry Name:CSK21_HUMAN