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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。
由于双链掺入法存在特异性较低的问题，1996年Heid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。
1Human Interleukin 12(IL-12/P70)
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.
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
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.
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.
This assay recognizes recombinant and natural human IL-12/P70. No
significant cross-reactivity or interference was observed.
The minimum detectable dose of human IL-12/P70 is typically less than 1.2
The sensitivity of this assay, or Lower Limit of Detection (LLD) was defined
as the lowest protein concentration that could be differentiated from zero.
Assay plate 1
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
1 x 20 ml
TMB Substrate 1 x 10 ml
Stop Solution 1 x 10 ml
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.
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
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.
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),
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
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.
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
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
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. 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
LIMITATIONS OF THE PROCEDURE
The kit should not be used beyond the expiration date on the kit label.
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.
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.
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
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
检测范围：4.7 pg/ml - 300 pg/ml
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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