價格詳細資料：試劑盒價格電議，齊一生物銷售：021－6034 8496；181214 53965；173021 04490,齊一生物科技（上海）有限公司提供ELISA試劑盒受到了廣大研究所.中科院科研單位一致肯定和認同。齊一生物大品牌保證，傾力為國內外科研院校實驗室提供*優質產品。

【兔血管內皮細胞粘附分子1(VCAM-1/CD106)ELISA試劑盒現貨】注意事項

1. 當混合蛋白溶液時應盡量輕緩，避免起泡。

2. 洗滌過程非常重要，不充分的洗滌易造成假陽性。

3. 一次加樣時間控制在5分鐘內，如標本數量多，推薦使用排槍加樣。

4. 請每次測定的同時做標準曲線，做復孔。

5. 如標本中待測物質含量過高，請先稀釋后再測定，計算時請*后乘以稀釋倍數。

6. 在配制標準品、檢測溶液工作液時，請以相應的稀釋液配制，不能混淆。

7. 底物請避光保存。

8. 不要用其它生產廠家的試劑替換試劑盒中的試劑。

FOR RESEARCH USE ONLY

Drug Names

Generic Name：

This kit can be used for determination of serum, plasma ａnd liquid samples Organization Content.

The experimental principle:

The product levels were measured in samples of the kit by double antibody sandwichmethod. The product with the purified antibody coated microtiter plate, made of solid phase antibody, to package is the product antigen monoclonal antibodies are then added to the micropores, the product and then with HRP labeled antibody binding, the formation of antibody - antigen - antibody complex enzyme label, after thorough washing with TMB chromogenic substrate. TMB in the HRP enzyme catalytic conversion into the blue, and in the action of acid into the final yellow. This product is positively related to the depth of color and in the samples. Instrument measured absorbance in the 450nm wavelength with ELISA (OD), the product concentration in the samples was calculated by standard curve.

Materials provided with the kit

Materials provided with the kit 96 determinations Storage

User manual   1  

Closure plate membrane   2  

Sealed bags   1  

Microelisa stripplate    1   2-8℃

Standard：360ng/L 0.5ml×1 bottle   2-8℃

Standard diluent  1.5ml×1 bottle   2-8℃

HRP-Conjugate reagent    6ml×1 bottle 2-8℃

Sample diluent    6ml×1 bottle 2-8℃

Chromogen Solution A 6ml×1 bottle 2-8℃

Chromogen Solution B 6ml×1 bottle 2-8℃

Stop Solution 6ml×1 bottle 2-8℃

wash  solution    （20ml×30 fold）

×1bottle  2-8℃

Specimen requirements

1.    serum- coagulation at room temperature 10-20 mins，centrifugation 20-min at the speed of 2000-3000 r.p.m. remove supernatant, If precipitation appeared, Centrifugal again. 【兔血管內皮細胞粘附分子1(VCAM-1/CD106)ELISA試劑盒現貨】

2.    plasma-use suited EDTA or citrate plasma as an anticoagulant,mix 10-20 mins ,centrifugation 20-min at the speed of 2000-3000 r.p.m. remove supernatant, If precipitation appeared, Centrifugal again.

3.    Urine-collect sue a sterile container, centrifugation 20-min at the speed of 2000-3000 r.p.m. remove supernatant, If precipitation appeared, Centrifugal again. The Operation of Hydrothorax ａnd cerebrospinal fluid Reference to it.

4.    cell culture supernatant-detect secretory components, collect sue a sterile container, centrifugation 20-min at the speed of 2000-3000 r.p.m. remove supernatant,detect the composition of cells, Dilut cell suspension with PBS（PH7.2-7.4）, Cell concentration reached 1 million / ml, repeated freeze-thaw cycles, damage cells ａnd release of intracellular components, centrifugation 20-min at the speed of 2000-3000 r.p.m. remove supernatant, If precipitation appeared, Centrifugal again.

5.    Tissue samples- After cutting samples, check the weight,add PBS（PH7.2-7.4）, Rapidly frozen with liquid nitrogen, maintain samples at 2-8℃ after melting,add PBS（PH7.4）, Homogenized by hand or Grinders, centrifugation 20-min at the speed of 2000-3000 r.p.m. remove supernatant.

6.    extract as soon as possible after Specimen collection,and according to the relevant literature, ａnd should be experiment as soon as possible after the extraction. If it can’t, specimen can be kept in -20 ℃ to preserve, Avoid repeated freeze-thaw cycles.

7.    Can’t detect the sample which contain NaN3, because NaN3 inhibits HRP active.

Assay procedure

1.Dilute ａnd add sample to Standard: set 10 Standard wells on the ELISA plates coated, add Standard 100μl to the first ａnd the second well, then add Standard dilution 50μl to the first ａnd the second well, mix; take out 100μl form the first ａnd the second well then add it to the third ａnd the forth well separately. then add Standard dilution 50μl to the third ａnd the forth well ,mix ; then take out 50μl from the third ａnd the forth well discard, add 50μl to the fifth ａnd the sixth well ,then add Standard dilution 50μl to the fifth ａnd the sixth well, mix ; take out 50μl from the fifth ａnd the sixth well ａnd add to the seventh ａnd the eighth well, then add Standard dilution 50μl to the seventh ａnd the eighth well ,mix ; take out 50μl from the seventh ａnd the eighth well ａnd add to the ninth ａnd the tenth well, add Standard dilution 50μl to the ninth ａnd the tenth well, mix , take out 50μl from the ninth ａnd the tenth well discard(add Sample 50μl to each well after Diluting ,(density: 240ng/L,160ng/L ,80ng/L,40ng/L, 20ng/L)

2.add sample：Set blank wells separately (blank comparison wells .don’t add sample ａnd HRP-Conjugate reagent, other each step operation is same). testing sample well. add Sample dilution 40μl to testing sample well, then add testing sample 10μl (sample final dilution is 5-fold), add sample to wells , don’t touch the well wall as far as possible, ａnd Gently mix. 【兔血管內皮細胞粘附分子1(VCAM-1/CD106)ELISA試劑盒現貨】

3.Incubate: After closing plate with Closure plate membrane ,incubate for 30 min at 37℃.

4.Configurate liquid: 30-fold (or 20-fold)wash solution diluted 30-fold (or 20-fold) with distilled water ａnd reserve.

5.washing：Uncover Closure plate membrane, discard Liquid, dry by swing, add washing buffer to every well, still for 30s then drain, repeat 5 times, dry by pat.

6.add enzyme：Add HRP-Conjugate reagent 50μl to each well, except  blank well.

7.incubate：Operation with 3.

8.washing：Operation with 5.

9.color：Add Chromogen Solution A 50ul ａnd Chromogen Solution B to each well, evade the light preservation for 15 min at 37℃

10.Stop the reaction：Add Stop Solution50μl to each well, Stop the reaction(the blue color change to yellow color).

11.assay：take blank well as zero , Read absorbance at 450nm after Adding Stop Solution ａnd within 15min.

Important notes

1.    The kit takes out from the refrigeration environment should be balanced 15-30 minutes in the room temperature, ELISA plates coated if has not use up after opened, the plate should be stored in Sealed bag.

2.    washing buffer will Crystallization separation, it can be heated the water helps dissolve when dilute . Washing does not affect the result.

3.    add Sample with sampler Each step, And proofread its accuracy frequently, avoids the experimental error. add sample within 5 mins, if the number of sample is much , recommend to use Volley . 【兔血管內皮細胞粘附分子1(VCAM-1/CD106)ELISA試劑盒現貨】

4.    if the testing material content is excessively higher (The sample OD is bigger than the first standard well ),please dilute Sample (n-fold), Please diluente ａnd multiplied by the dilution factor.（×n×5）.

5.    Closure plate membrane only limits the disposable use, to avoid cross-contamination.

6.    The substrate evade the light preservation.

7.    Please according to use instruction strictly, The test result determination must take the microtiter plate reader as a standard.

8.    All samples, washing buffer ａnd each kind of reject should according to infective material process.

9.    Do not mix reagents with those from other lots.

Calculate：

Take the standard density as the horizontal, the OD value for the vertical ,draw the standard curve on graph paper, Find out the corresponding density according to the sample OD value by the Sample curve, multiplied by the dilution multiple, or calculate the straight line regression equation of the standard curve with the standard density ａnd the OD value ,with the sample OD value in the equation, calculate the sample density, multiplied by the dilution factor, the result is the sample actual density.

Storage ａnd validity

1．Storage：  2-8℃.

2．validity： six months.

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QY-x1774 人CD4+T淋巴細胞         H9                                        

QY-x1775 胎兒骨髓間充質細胞（自建）     FBM                                     

QY-x1776 　     人腦細胞                                              

QY-x1777 人腦膠質細胞株     HEB                                      

QY-x1778 總T淋巴細胞          OKT3                                              【兔血管內皮細胞粘附分子1(VCAM-1/CD106)ELISA試劑盒現貨】

QY-x1779 BALB/C小鼠肝癌細胞    H22                                      

QY-x1780 小鼠前列腺癌細胞         RM-1                                             

QY-x1781 大鼠肝癌細胞（wistar 大鼠）     CBRH7919                                            

QY-x1782 小鼠肝癌細胞         HePa1-6                                       

QY-x1783 小鼠結腸癌     C26                                       

QY-x1784 穩定轉染了neor和LIF基因的STO細胞株 SNL                                       

QY-x1785 C57BL/6小鼠纖維肉瘤細胞   S180                                              

QY-x1786 小鼠艾氏腹水瘤細胞     ECA                                      

QY-x1787 大鼠腎上腺嗜鉻細胞瘤細胞         PC-12                                            

QY-x1788 大鼠膠質瘤     C6                                         

QY-x1789 褐鼠胰島素瘤上皮細胞         RIN-m                                           

QY-x1790 小鼠肥大細胞瘤細胞     P815                                              

QY-x1791 小鼠雜交瘤細胞     ST2/o                                             

QY-x1792 小鼠白血病細胞     P388                                              

QY-x1793 小鼠腎癌細胞         RenCa                                           

QY-x1794 小鼠胰島素瘤β細胞（NOD/Lt轉基因小鼠，SV40巨T抗原）       NIT-1                                              【兔血管內皮細胞粘附分子1(VCAM-1/CD106)ELISA試劑盒現貨】

QY-x1795 大鼠正常肝細胞     BRL                                      

QY-x1796 小鼠單核巨噬細胞         RAW 264.7                                            

QY-x1797 小鼠成纖維細胞     L929                                              

QY-x1798 BALBA小鼠成纖維細胞          BALB—3T3                                           

QY-x1799 早代小鼠胚胎成纖維細胞     MEF                                     

QY-x1800 中國倉鼠肺細胞     V79                                      

QY-x1801 非洲綠猴腎細胞     Vero                                     

QY-x1802 非洲綠猴腎細胞     Vero-E6                                        

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