Mouse GATA4 (GATA Binding Protein 4) ELISA Kit (MOES01063)

SKU:
MOES01063
Product type:
ELISA
Reactivity:
Mouse
ELISA Type:
Sandwich
€549

Description

ELISA Kit Technical ManualMSDS

Mouse GATA4 (GATA Binding Protein 4) 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 Mouse GATA4 . 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 Mouse GATA4 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 Mouse GATA4, 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 Mouse GATA4. The concentration of Mouse GATA4 in samples can be calculated by comparing the OD of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityMouse
Detection MethodColormetric
Detection Range0.31-20 ng/mL
Sensitivity0.19 ng/mL
Sample Volume Required Per Well100uL
Sample TypeSerum, plasma and other biological fluids

Specificity

This kit recognizes Mouse GATA4 in samples. No significant cross-reactivity or interference between Mouse GATA4 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
202.378
2.432
2.4052.335
101.507
1.525
1.5161.446
50.908
0.888
0.8980.828
2.50.439
0.441
0.440.37
1.250.244
0.224
0.2340.164
0.630.164
0.162
0.1630.093
0.310.109
0.127
0.1180.048
00.068
0.072
0.07--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse GATA4 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse GATA4 were tested on 3 different plates, 20 replicates in each plate.

Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (ng/mL)1.063.146.840.962.986.53
Standard deviation0.060.150.290.050.140.35
C V (%)5.664.784.245.214.705.36

Recovery

The recovery of Mouse GATA4 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)87-10294
EDTA plasma (n=5)85-9890
Cell culture media (n=5)87-10294

Linearity

Samples were spiked with high concentrations of Mouse GATA4 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 (%)94-10690-10599-114
Average (%)9997105
1:4Range (%)93-10685-9689-103
Average (%)999195
1:8Range (%)89-10282-9489-100
Average (%)948694
1:16Range (%)86-9880-9583-98
Average (%)938790

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

Mouse GATA4 (GATA Binding Protein 4) ELISA Kit (MOES01063) 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.

Mouse GATA4 (GATA Binding Protein 4) ELISA Kit (MOES01063) Protein Information

UniProt Protein Function:GATA4: a conserved and ubiquitous member of the GATA family of zinc-finger transcription factors. Members of this family recognize the GATA motif which is present in the promoters of many genes. This protein is thought to regulate genes involved in embryogenesis and in myocardial differentiation and function. May regulate a set of cardiac-specific genes and play a crucial role in cardiogenesis. Defects are a cause of atrial septal defect 2 (ASD2). ASD2 is an autosomal dominant condition with atrial septal defect and other congenital heart disease but no conduction defects or noncardiac abnormalities.
UniProt Protein Details:

Protein type:Motility/polarity/chemotaxis; Transcription factor

Cellular Component: nuclear chromatin; nucleus; transcription factor complex

Molecular Function:chromatin binding; DNA binding; metal ion binding; NFAT protein binding; protein binding; protein kinase binding; RNA polymerase II transcription factor activity, enhancer binding; sequence-specific DNA binding; transcription activator binding; transcription coactivator activity; transcription factor activity; transcription factor binding; zinc ion binding

Biological Process: BMP signaling pathway; cardiac muscle cell differentiation; cardiac muscle development; cardiac muscle hypertrophy; cell development; cell-cell signaling; embryonic digestive tract morphogenesis; embryonic foregut morphogenesis; embryonic heart tube anterior/posterior pattern formation; embryonic heart tube development; embryonic morphogenesis; endoderm formation; gastrulation with mouth forming second; heart development; heart looping; heart morphogenesis; in utero embryonic development; male gonad development; negative regulation of autophagy; positive regulation of angiogenesis; positive regulation of BMP signaling pathway; positive regulation of cardiac muscle cell proliferation; positive regulation of transcription from RNA polymerase II promoter; positive regulation of transcription, DNA-dependent; regulation of cardiac muscle cell proliferation; regulation of gene expression; regulation of transcription from RNA polymerase II promoter; regulation of transcription, DNA-dependent; response to drug; response to estrogen stimulus; response to mechanical stimulus; response to retinoic acid; Sertoli cell differentiation; spermatogenesis; transcription from RNA polymerase II promoter; transcription, DNA-dependent; tube morphogenesis; Wnt receptor signaling pathway through beta-catenin

UniProt Code:Q08369
NCBI GenInfo Identifier:46909569
NCBI Gene ID:14463
NCBI Accession:NP_032118.2
UniProt Secondary Accession:Q08369,P97491, Q9QZK4, B9EHF7,
UniProt Related Accession:Q08369
Molecular Weight:44,580 Da
NCBI Full Name:transcription factor GATA-4 isoform 2
NCBI Synonym Full Names:GATA binding protein 4
NCBI Official Symbol:Gata4
NCBI Official Synonym Symbols:Gata-4
NCBI Protein Information:transcription factor GATA-4
UniProt Protein Name:Transcription factor GATA-4
UniProt Synonym Protein Names:GATA-binding factor 4
Protein Family:Transcription factor
UniProt Gene Name:Gata4
UniProt Entry Name:GATA4_MOUSE
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