产品名称: 牛催乳素(PRL/LTH)ELISA Kit
英文名称: Bovine Prolactin/Luteotropic Hormone,PRL/LTH ELISA Kit
货号: CSB-E14002B
规格: 96T
种属: Bovine
待测物名称: prolactin
缩写: PRL
蛋白功能1: fertilization
蛋白功能3: Lactation
检测范围: 32 μIU/ml-8000 μIU/ml
灵敏度: 32 μIU/ml
反应时间: 1-5h
所需样本体积: 50-100ul
检测波长: 450 nm
用途: For research use only. Not for diagnostic use.
原理 : This assay employs the quantitative sandwich enzyme immunoassay technique. Antibody specific for PRL has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any PRL present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for PRL is added to the wells. After washing, avidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of PRL bound in the initial step. The color development is stopped and the intensity of the color is measured.
特异性: This assay has high sensitivity and excellent specificity for detection of Bovine PRL. No significant cross-reactivity or interference between Bovine PRL and analogues was observed.
精密度: Intra-assay Precision (Precision within an assay): CV%<8%
Three samples of known concentration were tested twenty times on one plate to assess.
Inter-assay Precision (Precision between assays): CV%<10%
Three samples of known concentration were tested in twenty assays to assess.
样本搜集及储存: Serum: Use a serum separator tube (SST) and allow samples to clot for two hours at room temperature or overnight at 4°C before centrifugation for 15 minutes at 1000 ×g. Remove serum and assay immediately or aliquot and store samples at -20°C or -80°C. Avoid repeated freeze-thaw cycles.
Plasma: Collect plasma using EDTA, or heparin as an anticoagulant. Centrifuge for 15 minutes at 1000 ×g at 2-8°C within 30 minutes of collection. Assay immediately or aliquot and store samples at -20°C or -80°C. Avoid repeated freeze-thaw cycles.
检测步骤: Bring all reagents and samples to room temperature before use. Centrifuge the sample again after thawing before the assay. It is recommended that all samples and standards be assayed in duplicate.
1. Prepare all reagents, working standards, and samples as directed in the previous sections.
2. Refer to the Assay Layout Sheet to determine the number of wells to be used and put any remaining wells and the desiccant back into the pouch and seal the ziploc, store unused wells at 4°C.
3. Add 100μl of standard and sample per well. Cover with the adhesive strip provided. Incubate for 2 hours at 37°C. A plate layout is provided to record standards and samples assayed.
4. Remove the liquid of each well, don't wash.
5. Add 100μl of Biotin-antibody (1x) to each well. Cover with a new adhesive strip. Incubate for 1 hour at 37°C. (Biotin-antibody (1x) may appear cloudy. Warm up to room temperature and mix gently until solution appears uniform.)
6. Aspirate each well and wash, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (200μl) using a squirt bottle, multi-channel pipette, manifold dispenser, or autowasher, and let it stand for 2 minutes, complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining wash Buffer by aspirating ordecanting. Invert the plate and blot it against clean paper towels.
7. Add 100μl of HRP-avidin (1x) to each well. Cover the microtiter plate with a new adhesive strip. Incubate for 1 hour at 37°C.
8. Repeat the aspiration/wash process for five times as in step 6.
9. Add 90μl of TMB Substrate to each well. Incubate for 15-30 minutes at 37°C. Protect from light.
10. Add 50μl of Stop Solution to each well, gently tap the plate to ensure thorough mixing.
11. Determine the optical density of each well within 5 minutes, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. Subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.
结果计算: Using the professional soft "Curve Expert 1.3" to make a standard curve is recommended, which can be downloaded from our web.
Average the duplicate readings for each standard 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 x-axis against the concentration on the y-axis and draw a best fit curve through the points on the graph. The data may be linearized by plotting the log of the PRL 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.
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蛋白
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Recombinant Cyprinus carpio Prolactin(prl) CSB-YP018724EQE 1mg Yeast
Recombinant Rat Prolactin(Prl) CSB-EP018724RA 1mg E.coli
Recombinant Rat Prolactin(Prl) CSB-YP018724RA 1mg Yeast
Recombinant Ictalurus punctatus Prolactin(prl) CSB-EP018724ILX 1mg E.coli
Recombinant Ictalurus punctatus Prolactin(prl) CSB-YP018724ILX 1mg Yeast
Recombinant Dicentrarchus labrax Prolactin(prl) CSB-EP018724DKC 1mg E.coli
Recombinant Dicentrarchus labrax Prolactin(prl) CSB-YP018724DKC 1mg Yeast
Recombinant Salmo salar Prolactin(prl) CSB-EP018724SWI 1mg E.coli
Recombinant Salmo salar Prolactin(prl) CSB-YP018724SWI 1mg Yeast
Recombinant Coregonus autumnalis Prolactin(prl) CSB-EP018724CPU 1mg E.coli
Recombinant Coregonus autumnalis Prolactin(prl) CSB-YP018724CPU 1mg Yeast
Recombinant Chelonia mydas Prolactin CSB-EP018724CHW 1mg E.coli
Recombinant Chelonia mydas Prolactin CSB-YP018724CHW 1mg Yeast
Recombinant Mesocricetus auratus Prolactin(PRL) CSB-EP018724MRG 1mg E.coli
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Recombinant Hypophthalmichthys nobilis Prolactin(prl) CSB-EP018724HRS 1mg E.coli
Recombinant Hypophthalmichthys nobilis Prolactin(prl) CSB-YP018724HRS 1mg Yeast
Recombinant Mustela vison Prolactin(PRL) CSB-EP018724MVE 1mg E.coli
Recombinant Mustela vison Prolactin(PRL) CSB-YP018724MVE 1mg Yeast
Recombinant Hypophthalmichthys molitrix Prolactin(prl) CSB-EP018724HRR 1mg E.coli
Recombinant Hypophthalmichthys molitrix Prolactin(prl) CSB-YP018724HRR 1mg Yeast
Recombinant Anguilla anguilla Prolactin(prl) CSB-EP018724BZF 1mg E.coli
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Recombinant Balaenoptera borealis Prolactin(PRL) CSB-EP018724BEA 1mg E.coli
Recombinant Balaenoptera borealis Prolactin(PRL) CSB-YP018724BEA 1mg Yeast
Recombinant Bufo japonicus Prolactin CSB-EP018724BNH 1mg E.coli
Recombinant Bufo japonicus Prolactin CSB-YP018724BNH 1mg Yeast
Recombinant Protopterus aethiopicus Prolactin(prl) CSB-EP018724PSV 1mg E.coli
Recombinant Protopterus aethiopicus Prolactin(prl) CSB-YP018724PSV 1mg Yeast
Recombinant Oncorhynchus mykiss Prolactin(prl) CSB-EP018724OEI 1mg E.coli
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Recombinant Camelus dromedarius Prolactin(PRL) CSB-EP018724CYV 1mg E.coli
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Recombinant Loxodonta africana Prolactin(PRL) CSB-EP018724LJC 1mg E.coli
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Recombinant Monodelphis domestica Prolactin(PRL) CSB-EP018724MUE 1mg E.coli
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Recombinant Sparus aurata Prolactin(prl) CSB-EP018724FJX 1mg E.coli
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Recombinant Trichosurus vulpecula Prolactin(PRL) CSB-EP018724TQG 1mg E.coli
Recombinant Trichosurus vulpecula Prolactin(PRL) CSB-YP018724TQG 1mg Yeast
Recombinant Sheep Prolactin(PRL) CSB-EP018724SH 1mg E.coli
Recombinant Sheep Prolactin(PRL) CSB-YP018724SH 1mg Yeast
Recombinant Pig Prolactin(PRL) CSB-EP018724PI 1mg E.coli
Recombinant Pig Prolactin(PRL) CSB-YP018724PI 1mg Yeast
Recombinant Mouse Prolactin(Prl) CSB-EP018724MO 1mg E.coli
Recombinant Mouse Prolactin(Prl) CSB-YP018724MO 1mg Yeast
Recombinant Human Prolactin(PRL) CSB-EP018724HU 1mg E.coli
Recombinant Human Prolactin(PRL) CSB-YP018724HU 1mg Yeast
Recombinant Bovine Prolactin(PRL) CSB-EP018724BO 1mg E.coli
Recombinant Bovine Prolactin(PRL) CSB-YP018724BO 1mg Yeast
Recombinant Macaca mulatta (Rhesus macaque) Prolactin(PRL) CSB-EP018724 1mg E.coli
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Recombinant Cat Prolactin(PRL) CSB-EP018724CA 1mg E.coli
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