產(chǎn)品名稱(chēng):Human IL-4 人白介4
產(chǎn)品型號:1600/48T; 2600/9
產(chǎn)品特點(diǎn):Human IL-4 人白介4定量檢測索萊寶ELISA試劑盒的優(yōu)勢:1,包被的酶標板單板可拆(能拆分成12個(gè)8孔的酶標條)2,提供免費代測代檢服務(wù),代出實(shí)驗數據3,發(fā)文章高獎勵4,磁鐵可吸附式包裝盒,包裝精美耐用
免費咨詢(xún):010-50973130
For research use only. Not for use in diagnostic procedures.
Human IL-4 人白介4?
MANUFACTURED AND DISTRIBUTED BY:
Country︱Company: China︱Beijing Solarbio Science & Technology Co., Ltd
Address: NO.8, Liandong U Valley, Tongzhou District, Beijing, P.R.China.
TABLE OF CONTENTS
SECTION PAGE
BACKGROUND...........................................................................................3
PRINCIPLE OF THE ASSAY......................................................................3
TECHNICAL HINTS AND LIMITATIONS...............................................4
PRECAUTIONS............................................................................................4
KIT COMPONENTS& STORAGE CONDITIONS.....................................5
OTHER SUPPLIES REQUIRED BUT NOT SUPPLIED............................6
SPECIMEN COLLECTION & STORAGE..................................................6
REAGENTS PREPARATION......................................................................6
ASSAY PROCEDURE .................................................................................8
CALCULATION OF RESULTS...................................................................8
PERFORMANCE CHARACTERISTICS....................................................10
REFERENCES..............................................................................................11
Human IL-4 人白介4?
BACKGROUND
The interleukin 4 (IL4, IL-4) is a cytokine that induces differentiation of naive helper T cells (Th0 cells) to Th2 cells. Upon activation by IL-4, Th2 cells subsequently produce additional IL-4 in a positive feedback loop. The cell that initially produces IL-4, thus inducing Th0 differentiation, has not been identified, but recent studies suggest that basophils may be the effector cell.It is closely related and has functions similar to Interleukin 13.IL-4 has been shown to drive mitogenesis, dedifferentiation, and metastasis in rhabdomyosarcoma.IL-4, along with other Th2 cytokines, is involved in the airway inflammation observed in the lungs of patients with allergic asthma.
PRINCIPLE OF THE ASSAY
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for IL-4 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IL-4 present is captured by the coated antibody after incubation. Following extensive washing, a biotin-conjugate antibody specific for IL-4 is added to detect the captured IL-4 protein in sample. For signal development, horseradish peroxidase (HRP)-conjugated Streptavidin is added, followed by Tetramethyl-benzidine (TMB) reagent. Following a wash to remove any unbound combination, and enzyme conjugate is added to the wells. Solution containing sulfuric acid is used to stop color development and the color intensity which is proportional to the quantity of bound protein is measurable at 450nm.
TECHNICAL HINTS AND LIMITATIONS
PRECAUTIONS
The Stop Solution suggested for use with this kit is an acid solution. Wear protective gloves, clothing, eye, and face protection. Wash hands thoroughly after handling.
KIT COMPONENTS& STORAGE CONDITIONS
PART | SIZE | STORAGE OF OPENED/ RECONSTITUTED MATERIAL |
Microwell Plate - antibody coated 96-well Microplate (8 wells ×12 strips) | 1 plate | Return unused wells to the foil pouch containing the desiccant pack. Reseal along entire edge of the zip-seal. May be stored for up to 1 month at 2 – 8℃** |
Standard - lyophilized,1000 pg/ml upon reconstitution | 2 vials | Aliquot and Store at -20°C** for six months |
Concentrated Biotin-Conjugated antibody(100X) - 120 ul/vial | 1 vial | Store at 2-8°C **for six months |
Concentrated Streptavidin-HRP solution(100X) - 120 ul/vial | 1 vial | Store at 2-8°C** for six months |
Standard /sample Diluent - 16 ml/vial | 1 bottle | Store at 2-8°C** for six months |
Biotin-Conjugate antibody Diluent - 16 ml/vial | 1 bottle | Store at 2-8°C** for six months |
Streptavidin-HRP Diluent - 16 ml/vial | 1 bottle | Store at 2-8°C** for six months |
Wash Buffer Concentrate (20x) - 30 ml/vial | 1 bottle | Store at 2-8°C** for six months |
Substrate Solution - 12 ml/vial | 1 bottle | Store at 2-8°C** for six months |
Stop Solution - 12 ml/vial | 1 bottle | Store at 2-8°C** for six months |
Plate Cover Seals | 4 pieces |
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**Provided this is within the expiration date of the kit.
OTHER SUPPLIES REQUIRED BUT NOT SUPPLIED
SPECIMEN COLLECTION & STORAGE
Cell Culture Supernates - Centrifuge cell culture media at 1000×g to remove debris. Assay immediay or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles.
Serum - Use a serum separator tube (SST) and allow samples to clot for 2 hours at room temperature or overnight at 2-8℃. Centrifuge at approximay for 15 minutes at 1000×g. Assay immediay or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles.
Plasma - Collect plasma using EDTA, heparin, or citrate as an anticoagulant. Centrifuge for 15 minutes at 1000×g within 30 minutes of collection. Assay immediay or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles.
Note: The normal human serum or plasma samples are suggested to make a 1:2 dilution.
REAGENTS PREPARATION
*If you do not run out of re-melting standard, store it at -20℃. Diluted standard shall not be reused.
*The working solution should be used within one day after dilution.
*The working solution should be used within one day after dilution.
Preparation of IL-4 standard dilutions
ASSAY PROCEDURE
CALCULATION OF RESULTS
Typical data using the IL-4 ELISA
Standard(pg/ml) | OD. | OD. | Average | Corrected |
0 | 0.044 | 0.04 | 0.042 | ------ |
3.12 | 0.097 | 0.092 | 0.0945 | 0.0525 |
6.25 | 0.13 | 0.106 | 0.118 | 0.076 |
12.5 | 0.229 | 0.233 | 0.231 | 0.189 |
25 | 0.42 | 0.412 | 0.416 | 0.374 |
50 | 0.741 | 0.734 | 0.7375 | 0.6955 |
100 | 1.315 | 1.306 | 1.3105 | 1.2685 |
200 | 2.205 | 2.212 | 2.2085 | 2.1665 |
Representative standard curve for IL-4 ELISA.
Performance Characteristics
SENSITIVITY: The minimum detectable dose was 1.5pg/mL.
SPECIFICITY: This assay recognizes both natural and recombinant human IL-4. The factors listed below were prepared at 100ng/ml in Standard /sample Diluent and assayed for cross-reactivity and no significant cross-reactivity or interference was observed.
Factors assayed for cross-reactivity
Recombinant human | Recombinant mouse | Recombinant porcine |
G-CSF | IL-1 | bovine FGF acidic |
GM-CSF | IL-4 | bovine FGF basic |
IL-1α_ | IL-3 | human PDGF |
IL-1β_ | IL-4 | porcine PDGF |
IL-2 sRα_ | IL-5 |
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IL-3 |
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IL-4 |
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IL-6 |
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IL-7 |
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IL-8 |
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LIF |
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TGF-β1 |
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TGF-β2 |
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REPEATABILITY: The coefficient of variation of both intra-assay and inter-assay were less than 10%.
RECOVERY:The recovery of IL-4 spiked to three different levels in four samples throughout the range of the assay in various matrices was evaluated.
Recovery of IL-4 in two matrices
Sample Type | Average % of Expected Range (%) | Range (%) |
Citrate plasma | 88 | 82-94 |
Cell culture supernatants | 97 | 92-102 |
LINEARITY:To assess the linearity of the assay, three samples were spiked with high concentrations of IL-4 in various matrices and diluted with the appropriate Sample Diluent to produce samples with values within the dynamic range of the assay.
The linearity of the assay
Dilution ratio | Recovery (%) | Citrate plasma | Cell culture supernatants |
1:2 | Average% of Expected | 85 | 96 |
Range (%) | 81-89 | 92-100 | |
1:4 | Average% of Expected | 92 | 102 |
Range (%) | 87-97 | 96-108 |
REFERENCES
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