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Human Adiponectin ELISA哪家好公司
发表时间:2017-10-14 08:27:00 文章来源:济源生物
Human Adiponectin ELISA
Human Adiponectin ELISA哪家好公司。伤寒杆菌有三种抗原:分别为菌体抗原(O抗原)、鞭毛抗原(H抗原)、体表抗原(Vi抗原)。其中以O抗原及H抗原的抗原性较强,而Vi抗原的抗原性不强,且相应抗体效价低且为时短暂,随细菌的消除而消失,故不列为肥达试验的检测项目。(但其检查有助于发现伤寒带菌者。)当抗原遇到特异性抗体(抗O及抗H)时,便会发生凝集反应,通过对凝集物量多少来推算病人体内抗体的多少,以协助诊断、治疗及判断预后。

广州市济源生物科技股份有限公司在经过多年市场化经营后,现在成为一家以Human Adiponectin ELISA、猴脂联素检测试剂盒、Monkey Adiponectin ELISA、人脂联素检测试剂盒为一体的综合性服务公司。济源生物始终坚持“以诚信为本、以质量为纲领”的服务宗旨,将以脂联素检测试剂盒产品技术优势、品质优势和服务优势转变成为用户的应用优势,使用户在Human Adiponectin ELISA哪家好市场竞争中永远处于领先地位。


基因兴奋剂大致有:(1)红细胞生成素的基因;(2)让肌肉纤维长粗的基因;(3)促进肌肉再生毛细血管的基因,让肌肉从毛细血管获得更多营养;(4)提高肌肉耐力的基因;(5)促进肌肉生长的基因;(6)抑制脂肪细胞的基因,可让人只长肉,不长膘;(7)抑制肌肉酸痛的基因。

据报道,这些基因兴奋剂用了没有,哪些用了,哪些没用,目前都还没有证据,但是必须防御。检测基因兴奋剂,就是检测人工注射的病毒和人工合成的基因,以及激素和酶。都能检测出来吗?还不知道。已知的,只有澳大利亚一个实验室可以检测人工合成的红细胞生成素的基因。

由于双链掺入法存在特异性较低的问题,1996年Heid[23]综合之前发现的Taq酶的5'核酸酶活性与荧光共振能量转移(fluorescenceresonance energy transfer,FRET)探针的概念提出了使用Taqman探针进行qPCR的方法。TaqMan探针的本质是FRET寡核苷酸探针,在探针的5'端标记荧光报告基团,3'端标记荧光淬灭基团,利用Taq酶具有5'3'外切酶活性,在PCR过程中水解与靶序列结合的寡核苷酸探针,使荧光基团得以游离,释放荧光信号。从而使能够与靶序列杂交的探针在扩增过程中释放荧光,通过real-timePCR的原理对其进行定量。由于其超高的特异性与成功的商品化推广,Taqman探针已经成为目前临床使用最为广泛的qPCR方法,其在各种病毒基因定量检测、基因分型、肿瘤相关基因表达检测等方面具有着不可替代的地位。

 1

Human Interleukin 12(IL-12/P70)

 

(96 tests)

 This immunoassay kit allows for the in vitro quantitative determination of human

IL-12/P70 concentrations in serum, plasma and other biological fluids.

 Expiration date six months from the date of manufacture

 FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.

2

INTRODUCTION

Interleukin 12 (IL-12) is an interleukin that is naturally produced by dendritic

cells, macrophages and human B-lymphoblastoid cells (NC-37) in response

to antigenic stimulation.

IL-12 is composed of a bundle of four alpha helices. It is a heterodimeric

cytokine encoded by two separate genes, IL-12A (p35) and IL-12B (p40).

The active heterodimer, and a homodimer of p40 are formed following

protein synthesis. IL-12 is involved in the differentiation of naive T cells into

Th1 cells, which is important in resistance against pathogens. It is known as

a T cell stimulating factor, which can stimulate the growth and function of T

cells. It stimulates the production of interferon-gamma (IFN-γ) and tumor

necrosis factor-alpha (TNF-α) from T and natural killer (NK) cells, and

reduces IL-4 mediated suppression of IFN-γ. T cells which produce IL-12

have a coreceptor, CD30, which is associated with IL-12 activity.

IL-12 plays an important role in the activities of natural killer cells and T

lymphocytes. IL-12 mediates enhancement of the cytotoxic activity of NK

cells and CD8+ cytotoxic T lymphocytes. There also seems to be a link

between IL-2 and the signal transduction of IL-12 in NK cells. IL-2

stimulates the expression of two IL-12 receptors, IL-12R-β1 and IL-12R-β2,

maintaining the expression of a critical protein involved in IL-12 signaling in

NK cells. Enhanced functional response is demonstrated by IFN-γ

production and killing of target cells.

IL-12 also has anti-angiogenic activity, which means it can block the

formation of new blood vessels. It does this by increasing production of

interferon gamma, which in turn increases the production of a chemokine

called inducible protein-10 (IP-10 or CXCL10). IP-10 then mediates this

3

anti-angiogenic effect. Because of its ability to induce immune responses

and its anti-angiogenic activity, there has been an interest in testing IL-12

as a possible anti-cancer drug. However, it has not been shown to have

substantial activity in the tumors tested to this date. There is a link that may

be useful in treatment between IL-12 and the diseases psoriasis and

inflammatory bowel disease.

PRINCIPLE OF THE ASSAY

The microtiter plate provided in this kit has been pre-coated with an

antibody specific to IL-12/P70. Standards or samples are then added to the

appropriate microtiter plate wells with a biotin-conjugated polyclonal

antibody preparation specific for IL-12/P70 and Avidin conjugated to

Horseradish Peroxidase (HRP) is added to each microplate well and

incubated. Then a TMB (3,3'5, 5' tetramethyl-benzidine) substrate solution

is added to each well. Only those wells that contain IL-12/P70,

biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a

change in color. The enzyme-substrate reaction is terminated by the

addition of a sulphuric acid solution and the color change is measured

spectrophotometrically at a wavelength of 450 nm ± 2 nm. The

concentration of IL-12/P70 in the samples is then determined by comparing

the O.D. of the samples to the standard curve.

DETECTION RANGE

4.7 pg/ml-300 pg/ml. The standard curve concentrations used for the

ELISA’s were 300 pg/ml, 150 pg/ml, 75 pg/ml, 37.5 pg/ml, 18.8 pg/ml, 9.4

pg/ml, 4.7 pg/ml.

4

SPECIFICITY

This assay recognizes recombinant and natural human IL-12/P70. No

significant cross-reactivity or interference was observed.

SENSITIVITY

The minimum detectable dose of human IL-12/P70 is typically less than 1.2

pg/ml.

The sensitivity of this assay, or Lower Limit of Detection (LLD) was defined

as the lowest protein concentration that could be differentiated from zero.

MATERIALS PROVIDED

Reagent Quantity

Assay plate 1

Standard 2

Sample Diluent 1 x 20 ml

Biotin-antibody Diluent 1 x 10 ml

HRP-avidin Diluent 1 x 10 ml

Biotin-antibody 1 x 120μl

HRP-avidin 1 x 120μl

Wash Buffer

1 x 20 ml

(25×concentrate)

TMB Substrate 1 x 10 ml

Stop Solution 1 x 10 ml

STORAGE

1. Unopened test kits should be stored at 2-8°C upon receipt and the

microtiter plate should be kept in a sealed bag. The test kit may be used

throughout the expiration date of the kit, provided it is stored as

prescribed above. Refer to the package label for the expiration date.

5

2. Opened test plate should be stored at 2-8°C in the aluminum foil bag

with desiccants to minimize exposure to damp air. The kits will remain

stable until the expiring date shown, provided it is stored as prescribed

above.

3. A microtiter plate reader with a bandwidth of 10 nm or less and an

optical density range of 0-3 OD or greater at 450nm wavelength is

acceptable for use in absorbance measurement.

REAGENT PREPARATION

Bring all reagents to room temperature before use.

1. Wash Buffer If crystals have formed in the concentrate, warm up to

room temperature and mix gently until the crystals have completely

dissolved. Dilute 20 ml of Wash Buffer Concentrate into deionized or

distilled water to prepare 500 ml of Wash Buffer.

2. Standard Centrifuge the standard vial at 6000-10000rpm for 30s.

Reconstitute the Standard with 1.0 ml of Sample Diluent. This

reconstitution produces a stock solution of 300 pg/ml.. Allow the

standard to sit for a minimum of 15 minutes with gentle agitation prior to

making serial dilutions. The undiluted standard serves as the high

standard (300 pg/ml.). The Sample Diluent serves as the zero

standard (0 pg/ml). Prepare fresh for each assay. Use within 4 hours

and discard after use.

3. Biotin-antibody Centrifuge the vial before opening. Dilute to the

working concentration using Biotin-antibody Diluent(1:100),

respectively.

6

4. HRP-avidin Centrifuge the vial before opening. Dilute to the working

concentration using HRP-avidin Diluent(1:100), respectively.

Precaution: The Stop Solution provided with this kit is an acid solution. Wear

eye, hand, face, and clothing protection when using this material.

OTHER SUPPLIES REQUIRED

 Microplate reader capable of measuring absorbance at 450 nm, with

the correction wavelength set at 540 nm or 570 nm.

 Pipettes and pipette tips.

 Deionized or distilled water.

 Squirt bottle, manifold dispenser, or automated microplate washer.

 An incubator which can provide stable incubation conditions up to

37°C±0.5°C.

SAMPLE COLLECTION AND STORAGE

 Serum Use a serum separator tube (SST) and allow samples to clot

for 30 minutes before centrifugation for 15 minutes at 1000 g. Remove

serum and assay immediately or aliquot and store samples at -20°C.

Centrifuge the sample again after thawing before the assay. Avoid

repeated freeze-thaw cycles.

 Plasma Collect plasma using citrate, EDTA, or heparin as an

anticoagulant. Centrifuge for 15 minutes at 1000 g within 30 minutes of

collection. Assay immediately or aliquot and store samples at -20°C.

Centrifuge the sample again after thawing before the assay. Avoid

repeated freeze-thaw cycles.

Note: Grossly hemolyzed samples are not suitable for use in this assay.

7

ASSAY PROCEDURE

Bring all reagents and samples to room temperature before use. It is

recommended that all samples, standards, and controls be assayed in duplicate.

All the reagents should be added directly to the liquid level in the well. The

pipette should avoid contacting the inner wall of the well.

1. Add 100μl of Standard, Blank, or Sample per well. Cover with the

adhesive strip. Incubate for 2 hours at 37°C.

2. Remove the liquid of each well, don’t wash.

3. Add 100μl of Biotin-antibody working solution to each well. Incubate

for 1 hour at 37°C. Biotin-antibody working solution may appear

cloudy. Warm up to room temperature and mix gently until solution

appears uniform.

4. Aspirate each well and wash, repeating the process three times for a

total of three washes. Wash: Fill each well with Wash Buffer (200μl) and

let it stand for 2 minutes, then remove the liquid by flicking the plate

over a sink. The remaining drops are removed by patting the plate on a

paper towel. Complete removal of liquid at each step is essential to

good performance.

5. Add 100μl of HRP-avidin working solution to each well. Cover the

microtiter plate with a new adhesive strip. Incubate for 1 hour at 37°C.

6. Repeat the aspiration and wash three times as step 4.

7. Add 90μl of TMB Substrate to each well. Incubate for 10-30 minutes at

37°C. Keeping the plate away from drafts and other temperature

fluctuations in the dark.

8

8. Add 50μl of Stop Solution to each well when the first four wells

containing the highest concentration of standards develop obvious blue

color. If color change does not appear uniform, gently tap the plate to

ensure thorough mixing.

9. Determine the optical density of each well within 30 minutes, using a

microplate reader set to 450 nm.

CALCULATION OF RESULTS

Using the professional soft "Curve Exert 1.3" to make a standard curve is

recommended, which can be downloaded from our web.

Average the duplicate readings for each standard, control, and sample and

subtract the average zero standard optical density. Create 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-12/P70 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.

LIMITATIONS OF THE PROCEDURE

 The kit should not be used beyond the expiration date on the kit label.

9

 Do not mix or substitute reagents with those from other lots or sources.

 It is important that the Standard Diluent selected for the standard curve

be consistent with the samples being assayed.

 If samples generate values higher than the highest standard, dilute the

samples with the appropriate Standard Diluent and repeat the assay.

 Any variation in Standard Diluent, operator, pipetting technique,

washing technique, incubation time or temperature, and kit age can

cause variation in binding.

 This assay is designed to eliminate interference by soluble receptors,

binding proteins, and other factors present in biological samples. Until

all factors have been tested in the Immunoassay, the possibility of

interference cannot be excluded.

TECHNICAL HINTS

 Centrifuge vials before opening to collect contents.

 When mixing or reconstituting protein solutions, always avoid foaming.

 To avoid cross-contamination, change pipette tips between additions of

each standard level, between sample additions, and between reagent

additions. Also, use separate reservoirs for each reagent.

 When using an automated plate washer, adding a 30 second soak

period following the addition of wash buffer, and/or rotating the plate

180 degrees between wash steps may improve assay precision.

 To ensure accurate results, proper adhesion of plate sealers during

incubation steps is necessary.

10

 Substrate Solution should remain colorless or light blue until added to

the plate. Keep Substrate Solution protected from light. Substrate

Solution should change from colorless or light blue to gradations of

blue.

 Stop Solution should be added to the plate in the same order as the

Substrate Solution. The color developed in the wells will turn from blue

to yellow upon addition of the Stop Solution. Wells that are green in

color indicate that the Stop Solution has not mixed thoroughly with the

Substrate Solution.

11

人白介素12(IL-12/P70)酶联免疫分析

试剂盒使用说明书

本试剂盒仅供研究使用

检测范围:4.7 pg/ml - 300 pg/ml

最低检测限:1.2 pg/ml

特异性:本试剂盒可同时检测天然或重组的人IL-12/P70,且与其他相关

蛋白无交叉反应。

有效期:6 个月

预期应用:ELISA法定量测定人血清、血浆、细胞培养上清或其它相关生

物液体中IL-12/P70 含量。

说明

1 试剂盒保存:未开封的试剂盒应储存于2-8℃;开封后的酶标板应与干燥

剂一起储存于铝箔袋中置于2-8℃保存。仅在此出储存条件下,产品在有

以上就是关于“Human Adiponectin ELISA的价格”的介绍。广州市济源生物科技股份有限公司是一家专业从事Human Adiponectin ELISA、猴脂联素检测试剂盒、猴脂联素检测试剂盒、Monkey Adiponectin ELISA的私营企业。济源生物相信,务实造就成功,创新成就未来。济源生物将一如既往地保持着创新的精神,不断以新的高品质的脂联素检测试剂盒产品服务于用户。如若您对该产品感兴趣的话,请拨打我们的热线:0755-84561234

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