91视频 - 8MAV,电影男人鸡巴插逼,亚洲 综合 图片视频,九九中文视频播放

產(chǎn)品詳情
  • 產(chǎn)品名稱:Gc ELISA Kit

  • 產(chǎn)品型號:
  • 產(chǎn)品廠商:國內(nèi)供應(yīng)3
  • 產(chǎn)品文檔:
你添加了1件商品 查看購物車
簡單介紹:
Gc ELISA Kit
詳情介紹:
Purpose This immunoassay kit allows for the in vitro quantitative determination of human DBP concentrations in cell culture supernates, serum, plasma and other biological fluids.
Sample Type Cell Culture Supernatant, Serum, Plasma, Biological Fluids
Analytical Method Quantitative
Detection Method Colorimetric
Specificity This assay recognizes recombinant and natural human DBP.
Cross-Reactivity (Details) No significant cross-reactivity or interference was observed.
Sensitivity <3.9 ng="" mlThe sensitivity of this assay, or Lower Limit of Detection (LLD) was defined as the lowest detectable concentration that could be differentiated from zero.
Characteristics Homo sapiens,Human,Vitamin D-binding protein,DBP,VDB,Gc-globulin,Group-specific component,GC
Components Reagent (Quantity): Assay plate (1×20ml), Standard (2), Sample Diluent (1×20ml), Assay Diluent A (1×10ml), Assay Diluent B (1×10ml), Detection Reagent A (1×120 μl), Detection Reagent B (1×120μl), Wash Buffer(25 x concentrate) (1×30ml), Substrate (1×10ml), Stop Solution (1×10ml), Plate sealer for 96 wells (5), Instruction (1)
Material not included Luminometer. Pipettes and pipette tips. EP tube Deionized or distilled water.
Background Also known as group specific protein (Gc), vitamin D binding protein (DBP) is a 52Kda protein that binds monomeric actin in addition to vitamin D. The protein is 458 residues in length, and forms three domains, the first of which contains the sterol . The three domains share limited sequence homology, with each other and to similar repeats in human serum albumin (HSA). The structure of DBP is also similar overall to HSA . It is presumed that the function of DBP binding actin is to clear up any actin that enters the blood stream as a result of cell injury especially crush injury to muscle. This function is also performed by gelsolin. The affinity for actin monomers is high (Kd = 10-9M) , and the actin has been reported to reside within domain III between residues 350 and 403, although the recent structural data of the complex shows that the interface is more extensive than this and involves residues from all three domains. The determination of the structure of the protein confirms the domain structure derived from the biochemical work. The structure of the complex of DBP and actin confirms that domain III forms an actin-binding contact, between sub-domains 1 and 3 of actin. However the extensive interface involves five distinct peptides from each of the three domains. Profilin and gelsolin compete for the DBP on G-actin, whereas DNase1 does not . By comparing the structure of free DBP and DBP:actin complex, it is argued there is only limited conformational change in VBDP upon binding actin .
Sample Volume 100 μL
Plate Pre-coated
Protocol The microtiter plate provided in this kit has been pre-coated with an antibody specific to DBP. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated polyclonal antibody preparation specific for DBP and Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB substrate solution is added to each well. Only those wells that contain DBP, 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 2 nm. The concentration of DBP in the samples is then determined by comparing the O.D. of the samples to the standard curve.
Reagent Preparation

Bring all reagents to room temperature before use. Wash Buffer - If crystals have formed in the concentrate, warm to room temperature and mix gently until the crystals have completely dissolved. Dilute 30 mL of Wash Buffer Concentrate into deionized or distilled water to prepare 750 mL of Wash Buffer. Standard - Reconstitute the Standard with 1.0 mL of Sample Diluent. This reconstitution produces a stock solution of 1,000 ng/ml . Allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making serial dilutions (Making serial dilution in the wells directly is not permitted). The undiluted standard serves as the high standard (1,00 0 ng/ml ). The Sample Diluent serves as the zero standard ( 0 ng/ml ). ng/mL 1,000 500 250 125 62.5 31.2 15.6 0 Detection Reagent A and B - Dilute to the working concentration using Assay Diluent A and B (1:100), respectively.

Sample Collection Serum - Use a serum separator tube (SST) and allow samples to clot for 30 minutes before centrifugation for 15 minutes at approximately 1000 × g. Remove serum and assay immediately or aliquot and store samples at -20C or -80C. Plasma - Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 1000 × g at 2 - 8C within 30 minutes of collection. Store samples at -20C or -80C. Avoid repeated freeze-thaw cycles. Cell culture supernates and other biological fluids - Remove particulates by centrifugation and assay immediately or aliquot and store samples at -20C or -80C. Avoid repeated freeze-thaw cycles. Note: Serum, plasma, and cell culture supernatant samples to be used within 7 days may be stored at 2-8 C, otherwise samples must stored at -20C (≤ 1 months) or -80C (≤ 2 months) to avoid loss of bioactivity and contamination. Avoid freeze-thaw cycles. When performing the assay slowly bring samples to room temperature.
Assay Procedure

Allow all reagents to reach room temperature (Please do not dissolve the reagents at 37C directly.). All the reagents should be mixed thoroughly by gently swirling before pipetting. Avoid foaming. Keep appropriate numbers of strips for 1 experiment and remove extra strips from microtiter plate. Removed strips should be resealed and stored at 4C until the kits expiry date. Prepare all reagents, working standards and samples as directed in the previous sections. Please predict the concentration before assaying. If values for these are not within the range of the standard curve, users must determine the optimal sample dilutions for their particular experiments.
1. Add 100 μl of Standard, Blank, or Sample per well. Cover with the Plate sealer. Incubate for 2 hours at 37C.
2. Remove the liquid of each well, don’t wash.
3. Add 100 μl of Detection Reagent A working solution to each well. Cover with the 4 Plate sealer. Incubate for 1 hour at 37C. Detection Reagent A working solution may appear cloudy. Warm 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 by filling each well with Wash Buffer (approximately 400 μl) using a squirt bottle, multi-channel pipette, manifold dispenser or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
5. Add 100 μl of Detection Reagent B working solution to each well. Cover with a new Plate sealer. Incubate for 1 hours at 37C.
6. Repeat the aspiration/wash as in step
4. 7. Add 90 μl of Substrate Solution to each well. Cover with a new Plate sealer. Incubate within 30 minutes at 37C. Protect from light.
8. Add 50 μl of Stop Solution to each well. If color change does not appear uniform, gently tap the plate to ensure thorough mixing.
9. Determine the optical density of each well at once, using a microplate reader set to 450 nm.
Important Note:
1. Absorbance is a function of the incubation time. Therefore, prior to starting the assay it is recommended that all reagents should be freshly prepared prior to use and all required strip-wells are secured in the microtiter frame. This will ensure equal elapsed time for each pipetting step, without interruption.
2. Please carefully reconstitute Standards or working Detection Reagent A and B according to the instruction, and avoid foaming and mix gently until the crystals have completely dissolved. The reconstituted Standards can be used only once. This assay requires pipetting of small volumes. To minimize imprecision caused by pipetting, ensure that pipettors are calibrated. It is recommended to suck more than 10μl for once pipetting.
3. To ensure accurate results, proper adhesion of plate sealers during incubation steps is necessary. Do not allow wells to sit uncovered for extended periods between incubation steps. Once reagents have been added to the well strips, DO NOT let the strips DRY at any time during the assay.
4. For each step in the procedure, total dispensing time for addition of reagents to the assay plate should not exceed 10 minutes.
5. 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.
6. The wash procedure is critical. Insufficient washing will result in poor precision and falsely elevated absorbance readings.
7. Duplication of all standards and specimens, although not required, is recommended.
8. Substrate Solution is easily contaminated. Please protect it from light. 5

Calculation of Results

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 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 DBP concentrations versus the log of the O.D. and the best fit line can be determined by regression analysis. It is recommended to use some related software to do this calculation, such as curve expert 13.0. 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.

Restrictions For Research Use only
Handling Advice 1. The kit should not be used beyond the expiration date on the kit label.
2. Do not mix or substitute reagents with those from other lots or sources.
3. If samples generate values higher than the highest standard, further dilute the 3 samples with the Assay 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.
4. This assay is designed to eliminate interference by soluble receptors, ligands, 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.
Storage 4 °C/-20 °C
Storage Comment The Standard, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20 °C upon being received. The other reagents can be stored at 4 °C.
日本 在线 内射| 亚洲国产精品色婷婷久久| 中午欧美日韩| 亚洲丝袜第一区| 亚洲欧美在线综合视频| 婷婷色情五月天电影| 国产日韩精品久久久| 中日韩中文字幕一区二区三| 日韩一区毛片| AV电影精品在线| 国产人与zoxxxx另类| 日韩欧美一区黄片| 台湾佬佬中文| 91夫妻福利视频| 小骚逼大片免费观看| av丝袜高清| 亚洲欧美中文字幕乱码在线| 69堂亚洲无码| 亚洲日韩中文字幕第一页| 有码在线观看完整版| av一区免费看不卡| 欧美性受ⅩXXX性爽| 操到疯狂乱喷av一区二区| 人人看人人爽人人澡| 婷婷一区二区三区四区中文字幕| 日韩一区二区三区欧洲在线观看| 日韩午夜av| 中文字幕日本3| 日韩成人电影一区2区3区| 国外网站AV一区二区三区四区 | 99re在线播放视频| 欧美午夜电影不卡在线观看| 精品 区一区二| 精品特级毛片网站| 一区二区yy中文字幕| 欧美日韩乱伦小说| 91麻豆精品久久毛片一级| 久久五码二区| 久久夜色国产精品噜噜AVAV| 美女白虎屌| 国产伊人精品大香蕉|