歡迎來北京茄子视频官网地址入口科技有限公司!茄子视频官网地址入口將為您提供周到的服務!
首頁 > 產品中心 > ELISA試劑盒 > 人 Human ELISA > Human IL-4 人白介4

產品名稱:Human IL-4 人白介4

產品型號:1600/48T; 2600/9

產品報價:

產品特點:Human IL-4 人白介4定量檢測
茄子视频官网下载地址ELISA試劑盒的優勢:
1,包被的酶標板單板可拆(能拆分成12個8孔的酶標條)
2,提供免費代測代檢服務,代出實驗數據
3,發文章高獎勵
4,磁鐵可吸附式包裝盒,包裝精美耐用

免費谘詢:010-50973132

發郵件給茄子视频官网地址入口:837939727@qq.com

分享到:

1600/48T; 2600/9Human IL-4 人白介4的詳細資料:

 For research use only. Not for use in diagnostic procedures.

Human IL-4 人白介4

MANUFACTURED AND DISTRIBUTED BY:

CountryCompany: ChinaBeijing 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

  • This Solarbio ELISA should not be used beyond the expiration data on the kit label.
  • To avoid cross-contamination, use a fresh reagent reservoir and pipette tips for each step.
  • To ensure accurate results, some details, such as technique, plasticware and water sources should be emphasized.
  • A thorough and consistent wash technique is essential for proper assay performance.
  • A standard curve should be generated for each set of samples assayed. 
  • It is recommended that all standards and samples be assayed in duplicate.
  • Avoid microbial contamination of reagents and buffers. Buffers containing protein should be made under aseptic conditions and be prepared fresh daily.
  • In order to ensure the accuracy of the results, the standard curve should be made every time.

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

 

 

**Provided this is within the expiration date of the kit.

OTHER SUPPLIES REQUIRED BUT NOT SUPPLIED

  • Microplate reader capable of measuring absorbance at 450 nm.
  • Pipettes and pipette tips.
  • Deionized or distilled water.
  • Squirt bottle, manifold dispenser, or automated microplate washer.
  • 500 mL graduated cylinder.
  • Human IL-4 controls (optional; available from Solarbio).

 

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

  • Temperature returning - Bring all kit components and specimen to room temperature (20-25) before use.
  • Wash Buffer - Dilute 30mL of Wash Buffer Concentrate with 570mL of deionized or distilled water to prepare 600mL of Wash Buffer. If crystals have formed in the concentrate Wash Buffer, warm to room temperature and mix gently until the crystals have compley dissolved.
  • StandardSpecimen - Reconstitute the Standard with 1.0mL of deionized or distilled water. This reconstitution produces a stock solution of 1000pg/mL. Allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making dilutions. Pipette 800mL of Standard/Specimen Diluent into the 200 pg/mL tube, and add 200mL stock solution of 1000 pg/mL into it to get the high standard of 200 pg/mL. Pipette 500mL of Standard/Specimen Diluent into the remaining tubes. Use the high standard to produce a 2-fold dilution series (below).. Mix each tube thoroughly and change pipette tips between each transfer. The 200pg/mL standard serves as the high standard. The Standard/specimen Diluent serves as the zero standard (0pg/mL).

*If you do not run out of re-melting standard, store it at -20. Diluted standard shall not be reused.

  • Working solution of Biotin-Conjugate anti-human IL-4 antibody: Make a 1:100 dilution of the concentrated Biotin-Conjugate solution with the Biotin-Conjugate antibody Diluent in a clean plastic tube.

*The working solution should be used within one day after dilution.

  • Working solution of Streptavidin-HRP: Make a 1:100 dilution of the concentrated Streptavidin-HRP solution with the Streptavidin-HRP Diluent in a clean plastic tube. 

*The working solution should be used within one day after dilution.

 

Preparation of IL-4 standard dilutions

 

ASSAY PROCEDURE

 CALCULATION OF RESULTS

  • The standard curve is used to determine the amount of specimens.
  • First, average the duplicate readings for each standard, control, and sample. All O.D. values are subtracted by the mean value of blank control before result interpretation.
  • Construct a standard curve by reducing the data using computer software capable of generating a four parameter logistic (4-PL) curve-fit. As an alternative, construct a standard curve by plotting the mean absorbance for each standard on the y-axis against the concentration on the x-axis and draw a best fit curve through the points on the graph.
  • The data may be linearized by plotting the log of the IL-4 concentrations versus the log of the O.D. and the best fit line can be determined by regression analysis. This procedure will produce an adequate but less precise fit of the data. If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor.
  • This standard curve is provided for demonstration only. A standard curve should be generated for each set of samples assayed.

 

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

 

IL-3

 

 

IL-4

 

 

IL-6

 

 

IL-7

 

 

IL-8

 

 

LIF

 

 

TGF-β1

 

 

TGF-β2

 

 

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 evalsuated.

 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

  • Sokol, C.L., Barton, G.M., Farr, A.G. & Medzhitov, R. (2008). "A mechanism for the initiation of allergen-induced T helper type 2 responses". Nat Immunol 9 (3): 310–318.
  • Liang, H-E, et al. (2012) Divergent expression patterns of IL-4 and IL-13 define unique functions in allergic immunity. Nature Immunology, 13: 58–66.
  • Howard M, Paul WE (1982). "Interleukins for B lymphocytes". Lymphokine Res. 1 (1): 1–4.
  • Yokota T et al. (1986). "Isolation and characterization of a human interleukin cDNA clone, homologous to mouse B-cell stimulatory factor 1, that express

 

 

 

 

 

 相關同類產品:

留言框

  • 產品:

  • 留言內容:

  • 您的單位:

  • 您的姓名:

  • 聯係電話:

  • 常用郵箱:

  • 詳細地址:

  • 省份:

  • 驗證碼:

    請輸入計算結果(填寫阿拉伯數字),如:三加四=7

版權所有  ICP備案號:京ICP備12051307號-5 管理登陸 技術支持:化工儀器網   總流量:252632  網站地圖

在線客服
在線客服
用心服務 成就你我