Description
Mouse GATA3 (GATA Binding Protein 3) ELISA Kit
The Mouse GATA3 (GATA Binding Protein 3) ELISA Kit is specifically designed for the accurate quantification of GATA3 levels in mouse serum, plasma, and cell culture supernatants. This kit boasts high sensitivity and specificity, guaranteeing precise and consistent results for various research purposes.GATA3 is a critical transcription factor involved in the regulation of gene expression, particularly in immune cells and the development of T-helper 2 (Th2) cells. Dysregulation of GATA3 levels has been implicated in various diseases, including asthma, allergic reactions, and certain types of cancer, making it a valuable biomarker for studying these conditions and exploring potential therapeutic interventions.
By using the Mouse GATA3 ELISA Kit, researchers can gain valuable insights into the role of GATA3 in immune responses, inflammation, and disease progression, ultimately advancing our understanding of these complex biological processes.
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Mouse |
| Detection Method: | Colormetric |
| Detection Range: | 15.63-1000 pg/mL |
| Sensitivity: | 9.38 pg/mL |
| Sample Volume Required Per Well: | 100µL |
| Sample Type: | Serum, plasma and other biological fluids |
| Specificity: | This kit recognizes Mouse GATA3 in samples. No significant cross-reactivity or interference between Mouse GATA3 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 Mouse GATA3. 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 GATA3 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 GATA3, 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 GATA3. The concentration of Mouse GATA3 in samples can be calculated by comparing the OD of the samples to the standard curve.
| UniProt Protein Function: | GATA3: a transcriptional regulator central in Th2 cell differentiation. Inhibits breast cancer growth and pulmonary breast cancer metastasis. Represses INK4C transcription, constrains luminal progenitor cell expansion, and suppresses luminal tumorigenesis in the mammary gland. High GATA3 and low INK4C expression predicts a favorable patient outcome in luminal A type breast tumors. IFN-lambda1 (IL-29) inhibits GATA3 expression and suppresses Th2 responses in human T cells. GATA3 haploinsufficiency leads to HDR (hypoparathyroidism, deafness, and renal dysplasia) syndrome. Two alternatively spliced human isoforms have been described. |
| UniProt Protein Details: | Protein type:Transcription factor Cellular Component: nuclear chromatin; nucleolus; nucleus; transcription factor complex Molecular Function:chromatin binding; DNA binding; interleukin-2 receptor binding; metal ion binding; protein binding; protein dimerization activity; sequence-specific DNA binding; transcription coactivator activity; transcription factor activity; transcription factor binding; zinc ion binding Biological Process: anatomical structure formation; axon guidance; cell activation; cell fate determination; cell maturation; cell morphogenesis; chromatin remodeling; developmental growth; ear development; embryonic hemopoiesis; embryonic organ development; erythrocyte differentiation; gut development; homeostasis of number of cells; humoral immune response; immune system process; in utero embryonic development; innate immune response; inner ear morphogenesis; kidney development; lens development in camera-type eye; male gonad development; mesonephros development; negative regulation of cell cycle; negative regulation of cell proliferation; negative regulation of fat cell differentiation; negative regulation of inflammatory response; negative regulation of interferon-gamma production; negative regulation of interleukin-2 production; negative regulation of mammary gland epithelial cell proliferation; negative regulation of transcription from RNA polymerase II promoter; negative regulation of transcription, DNA-dependent; nervous system development; neuron differentiation; neuron migration; norepinephrine biosynthetic process; parathyroid gland development; pharyngeal system development; phosphoinositide 3-kinase cascade; positive regulation of cell differentiation; positive regulation of cytokine production; positive regulation of interleukin-13 production; positive regulation of interleukin-4 production; positive regulation of interleukin-5 production; positive regulation of protein kinase B signaling cascade; positive regulation of signal transduction; positive regulation of T cell differentiation; positive regulation of transcription from RNA polymerase II promoter; positive regulation of transcription, DNA-dependent; post-embryonic development; pro-T cell differentiation; regulation of CD4-positive, alpha beta T cell differentiation; regulation of cytokine biosynthetic process; regulation of histone H3-K4 methylation; regulation of neuron apoptosis; regulation of transcription from RNA polymerase II promoter; regulation of transcription, DNA-dependent; signal transduction; sympathetic nervous system development; T cell differentiation; T cell differentiation in the thymus; T cell receptor signaling pathway; T-helper 2 cell differentiation; thymic T cell selection; thymus development; TOR signaling pathway; transcription, DNA-dependent; uterus development |
| UniProt Code: | P23772 |
| NCBI GenInfo Identifier: | 6679951 |
| NCBI Gene ID: | 14462 |
| NCBI Accession: | NP_032117. 1 |
| UniProt Related Accession: | P23772 |
| Molecular Weight: | 47,968 Da |
| NCBI Full Name: | trans-acting T-cell-specific transcription factor GATA-3 |
| NCBI Synonym Full Names: | GATA binding protein 3 |
| NCBI Official Symbol: | Gata3 |
| NCBI Official Synonym Symbols: | jal; Gata-3 |
| NCBI Protein Information: | trans-acting T-cell-specific transcription factor GATA-3 |
| UniProt Protein Name: | Trans-acting T-cell-specific transcription factor GATA-3 |
| UniProt Synonym Protein Names: | GATA-binding factor 3 |
| Protein Family: | Transcription factor |
| UniProt Gene Name: | Gata3 |
| UniProt Entry Name: | GATA3_MOUSE |
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 (pg/mL) | O.D | Average | Corrected |
| 1000 | 2.4 2.434 | 2.417 | 2.351 |
| 500 | 1.658 1.698 | 1.678 | 1.612 |
| 250 | 1.005 0.973 | 0.989 | 0.923 |
| 125 | 0.416 0.442 | 0.429 | 0.363 |
| 62.5 | 0.282 0.266 | 0.274 | 0.208 |
| 31.25 | 0.169 0.167 | 0.168 | 0.102 |
| 15.63 | 0.117 0.121 | 0.119 | 0.053 |
| 0 | 0.058 0.074 | 0.066 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse GATA3 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse GATA3 were tested on 3 different plates, 20 replicates in each plate.
| Intra-assay Precision | Inter-assay Precision | |||||
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 48.34 | 88.02 | 389.35 | 45.67 | 79.83 | 360.21 |
| Standard deviation | 3.38 | 4.44 | 14.48 | 2.37 | 3.76 | 11.67 |
| C V (%) | 6.99 | 5.04 | 3.72 | 5.19 | 4.71 | 3.24 |
Recovery
The recovery of Mouse GATA3 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) | 89-103 | 96 |
| EDTA plasma (n=5) | 92-108 | 99 |
| Cell culture media (n=5) | 87-102 | 93 |
Linearity
Samples were spiked with high concentrations of Mouse GATA3 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) | ||
| 1:2 | Range (%) | 96-110 | 97-113 | 95-108 |
| Average (%) | 104 | 104 | 102 | |
| 1:4 | Range (%) | 91-107 | 86-101 | 83-93 |
| Average (%) | 99 | 92 | 88 | |
| 1:8 | Range (%) | 89-100 | 79-90 | 87-102 |
| Average (%) | 94 | 85 | 93 | |
| 1:16 | Range (%) | 87-100 | 83-96 | 87-98 |
| Average (%) | 94 | 88 | 92 |
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 ×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 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 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.
- 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.
