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Human IL-2 ELISA-人白細胞介素2(IL-2)ELISA
  • Human IL-2 ELISA-人白細胞介素2(IL-2)ELISA
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貨物所在地:北京北京市

地: 北京市通州區馬駒橋鎮聯東U谷景

更新時間:2025-02-21 21:00:07

瀏覽次數:1085

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可用于檢測人血清/血漿/細胞培養上清中白細胞介素2的含量,達到定量分析,從而檢測其中的含量,用于后續研究和科學分析。
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中文名稱人白細胞介素2檢測試劑盒
英文名稱Human IL-2 ELISA KIT
ReactivityHuman
Sample typeSerum, plasma, Cell culture supernatant
Sensitivity7 pg/ml
Detection TargetIL-2
ApplicationsSandwich ELISA

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

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

BACKGROUND

Interleukin-2 (IL-2) is an interleukin, a type of cytokine signaling molecule in the immune system. It is a protein that regulates the activities of white blood cells (leukocytes, often lymphocytes) that are responsible for immunity. IL-2 is part of the body's natural response to microbial infection, and in discriminating between foreign ("non-self") and "self". IL-2 mediates its effects by binding to IL-2 receptors, which are expressed by lymphocytes. While the causes of itchiness are poorly understood, some evidence indicates that IL-2 is involved in itchy psoriasis.

 

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-Conjugateantibody(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 - 1ml/vial

1 bottle

Store at 2-8°C** for six months

Stop Solution - 1ml/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-2 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.
  • Standard\Specimen - 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 500mL of Standard/Specimen Diluent into 500pg/ml tube and the remaining tubes. Use the stock solution of 1000pg/mL to produce a 2-fold dilution series (below). Mix each tube thoroughly and change pipette tips between each transfer. The 500pg/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-2 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-2 standard dilutions

 

ASSAY PROCEDURE

Typical data using the IL-2 ELISA

Standard(pg/ml)

OD.

OD.

Average

Corrected

0

0.014

0.019

0.016

-------

7.81

0.090

0.082

0.086

0.069

15.62

0.148

0.142

0.145

0.128

31.25

0.250

0.258

0.254

0.237

62.5

0.519

0.593

0.556

0.539

125

0.943

1.050

0.996

0.980

250

1.942

1.964

1.953

1.936

500

3.091

3.136

3.113

3.096

 

 

 

Representative standard curve for IL-2 ELISA.

Performance Characteristics

SENSITIVITY: The minimum detectable dose was 4pg/mL.

SPECIFICITY: This assay recognizes both natural and recombinant human IL-2. 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

IL-2 sRa_

IL-2

bovine FGF acidic

IL-2 Ra_

IL-1

bovine FGF basic

IL-2 Rg_

IL-2

human PDGF

IL-3

IL-3

porcine PDGF

IL-4

IL-4

 

IL-6

IL-5

 

IL-7

IL-6

 

IL-8

IL-7

 

LIF

 

 

TGF-b1

 

 

TGF-b2

 

 

 TNF-a_

 

 

 TNF-b_

 

 

 

REPEATABILITY: The coefficient of variation of both intra-assay and inter-assay were less than 10%.

RECOVERY:The recovery of IL-2 spiked to three different levels in four samples throughout the range of the assay in various matrices was evaluated.

 

 Recovery of IL-2 in two matrices  

Sample Type

Average % of Expected Range (%)

Range (%)

Citrate plasma

86

82-91

Cell culture supernatants

95

90-100

LINEARITY:To assess the linearity of the assay, three samples were spiked with high concentrations of IL-2 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

91

94

Range (%)

85-97

86-103

1:4

Average% of Expected

98

96

Range (%)

89-108

90-103

 

 

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