Human GRIK2 (Glutamate Receptor, Ionotropic, Kainate 2) ELISA Kit (HUES02441)

SKU:
HUES02441
€539

Description

ELISA Kit Technical ManualMSDS

Human GRIK2 (Glutamate Receptor, Ionotropic, Kainate 2) 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 GRIK2 . 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 GRIK2 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 GRIK2, 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 GRIK2. The concentration of Human GRIK2 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 GRIK2 in samples. No significant cross-reactivity or interference between Human GRIK2 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
102.296
2.344
2.322.24
51.597
1.633
1.6151.535
2.50.955
0.937
0.9460.866
1.250.422
0.45
0.4360.356
0.630.295
0.271
0.2830.203
0.320.195
0.171
0.1830.103
0.160.132
0.134
0.1330.053
00.076
0.084
0.08--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human GRIK2 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human GRIK2 were tested on 3 different plates, 20 replicates in each plate.

Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (ng/mL)0.531.304.970.541.275.46
Standard deviation0.030.080.150.030.070.18
C V (%)5.666.153.025.565.513.30

Recovery

The recovery of Human GRIK2 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)94-108101
EDTA plasma (n=5)88-9893
Cell culture media (n=5)93-10599

Linearity

Samples were spiked with high concentrations of Human GRIK2 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 (%)99-11393-10790-104
Average (%)1059996
1:4Range (%)90-10383-9481-95
Average (%)958988
1:8Range (%)91-10581-9186-100
Average (%)978692
1:16Range (%)88-10180-9486-98
Average (%)958693

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 GRIK2 (Glutamate Receptor, Ionotropic, Kainate 2) ELISA Kit (HUES02441) 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 GRIK2 (Glutamate Receptor, Ionotropic, Kainate 2) ELISA Kit (HUES02441) Protein Information

UniProt Protein Function:GluR6: Ionotropic glutamate receptor. L-glutamate acts as an excitatory neurotransmitter at many synapses in the central nervous system. Binding of the excitatory neurotransmitter L- glutamate induces a conformation change, leading to the opening of the cation channel, and thereby converts the chemical signal to an electrical impulse. The receptor then desensitizes rapidly and enters a transient inactive state, characterized by the presence of bound agonist. May be involved in the transmission of light information from the retina to the hypothalamus. Modulates cell surface expression of NETO2. Defects in GRIK2 are the cause of mental retardation autosomal recessive type 6 (MRT6). It is characterized by significantly sub-average general intellectual functioning associated with impairments in adaptative behavior and manifested during the developmental period. In contrast to syndromic or specific mental retardation which also present with associated physical, neurological and/or psychiatric manifestations, intellectual deficiency is the only primary symptom of non-syndromic mental retardation. MRT6 patients display mild to severe mental retardation and psychomotor development delay in early childhood. Patients do not have neurologic problems, congenital malformations, or facial dysmorphism. Body height, weight, and head circumference are normal. Belongs to the glutamate-gated ion channel (TC 1.A.10.1) family. GRIK2 subfamily. 7 isoforms of the human protein are produced by alternative splicing.
UniProt Protein Details:

Protein type:Membrane protein, integral; Channel, calcium; Channel, ligand-gated; Membrane protein, multi-pass

Chromosomal Location of Human Ortholog: 6q16.3

Cellular Component: dendrite cytoplasm; presynaptic membrane; postsynaptic membrane; kainate selective glutamate receptor complex; integral to plasma membrane; dendrite; postsynaptic density; plasma membrane; terminal button; perikaryon; cell junction

Molecular Function:protein homodimerization activity; kainate selective glutamate receptor activity; extracellular-glutamate-gated ion channel activity; ubiquitin conjugating enzyme binding; ubiquitin protein ligase binding; PDZ domain binding

Biological Process: regulation of synaptic transmission; regulation of long-term neuronal synaptic plasticity; synaptic transmission, glutamatergic; behavioral fear response; glutamate signaling pathway; generation of action potential; negative regulation of synaptic transmission, glutamatergic; regulation of short-term neuronal synaptic plasticity; regulation of JNK cascade; receptor clustering; positive regulation of synaptic transmission; cellular calcium ion homeostasis; synaptic transmission; intracellular protein transport; regulation of inhibitory postsynaptic membrane potential; neuron apoptosis; ionotropic glutamate receptor signaling pathway; transport; positive regulation of neuron apoptosis; negative regulation of neuron apoptosis; regulation of excitatory postsynaptic membrane potential

Disease: Mental Retardation, Autosomal Recessive 6

NCBI Summary:Glutamate receptors are the predominant excitatory neurotransmitter receptors in the mammalian brain and are activated in a variety of normal neurophysiologic processes. This gene product belongs to the kainate family of glutamate receptors, which are composed of four subunits and function as ligand-activated ion channels. The subunit encoded by this gene is subject to RNA editing at multiple sites within the first and second transmembrane domains, which is thought to alter the structure and function of the receptor complex. Alternatively spliced transcript variants encoding different isoforms have also been described for this gene. Mutations in this gene have been associated with autosomal recessive mental retardation. [provided by RefSeq, Jul 2008]
UniProt Code:Q13002
NCBI GenInfo Identifier:2492627
NCBI Gene ID:2898
NCBI Accession:Q13002.1
UniProt Secondary Accession:Q13002,Q8WWS1, Q96KS6, Q96KS7, Q96KS8, A6NMY9, B5MCV0 D7RWZ3, D7RWZ4, D7RWZ5, D7RWZ6, D7RWZ7,
UniProt Related Accession:Q13002
Molecular Weight:908
NCBI Full Name:Glutamate receptor ionotropic, kainate 2
NCBI Synonym Full Names:glutamate receptor, ionotropic, kainate 2
NCBI Official Symbol:GRIK2
NCBI Official Synonym Symbols:EAA4; GLR6; MRT6; GLUK6; GLUR6; GluK2
NCBI Protein Information:glutamate receptor ionotropic, kainate 2; gluR-6; bA487F5.1; glutamate receptor 6; glutamate receptor form A; glutamate receptor form B; glutamate receptor form C; glutamate receptor form D; glutamate receptor form E; excitatory amino acid receptor 4
UniProt Protein Name:Glutamate receptor ionotropic, kainate 2
UniProt Synonym Protein Names:Excitatory amino acid receptor 4; EAA4; Glutamate receptor 6; GluR-6; GluR6
Protein Family:Glutamate receptor ionotropic
UniProt Gene Name:GRIK2
UniProt Entry Name:GRIK2_HUMAN
View AllClose

Additional Information

Product type:
ELISA
Reactivity:
Human
ELISA Type:
Sandwich
View AllClose