E(IgE)ELISA試劑盒FOR RESEARCH USE ONLY
Drug Names
Generic Name：E(IgE)ELISA試劑盒
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 kitE(IgE)ELISA試劑盒
Materials provided with the kit    48determinations    96 determinations    Storage
User manual    1    1    
Closure plate membrane    2    2    
Sealed bags    1    1    
Microelisa stripplate    1    1    2-8℃
Standard：360ng/L    0.5ml×1 bottle    0.5ml×1 bottle    2-8℃
Standard diluent    1.5ml×1 bottle    1.5ml×1 bottle    2-8℃
HRP-Conjugate reagent    3ml×1 bottle    6ml×1 bottle    2-8℃
Sample diluent    3ml×1 bottle    6ml×1 bottle    2-8℃
Chromogen Solution A    3ml×1 bottle    6ml×1 bottle    2-8℃
Chromogen Solution B    3ml×1 bottle    6ml×1 bottle    2-8℃
Stop Solution    3ml×1 bottle    6ml×1 bottle    2-8℃
wash  solution    （20ml×20 fold）
×1bottle    （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.
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 separay. 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 separay (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.
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 .
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.

代理品牌產(chǎn)品Cystatin C Monoclonal Antibody    YM0179    美國Immunoway    50ug, 100ug    WB ELISA    Human
Cytokeratin 15 Monoclonal Antibody    YM0180    美國Immunoway    50ug, 100ug    WB IHC IF FCM ELISA    Human
Cytokeratin 17 Monoclonal Antibody    YM0181    美國Immunoway    50ug, 100ug    WB IHC ELISA    Human
Cytokeratin 18 Monoclonal Antibody    YM0182    美國Immunoway    50ug, 100ug    WB IHC ELISA    Human
Cytokeratin 19 Monoclonal Antibody    YM0183    美國Immunoway    50ug, 100ug    WB IHC ELISA    Human
Cytokeratin 19 Monoclonal Antibody    YM0184    美國Immunoway    50ug, 100ug    WB ELISA    Human
Cytokeratin 19 Monoclonal Antibody    YM0185    美國Immunoway    50ug, 100ug    WB IHC ELISA    Human
Cytokeratin 19 Monoclonal Antibody    YM0186    美國Immunoway    50ug, 100ug    WB IHC IF ELISA    Human
Cytokeratin 5 Monoclonal Antibody    YM0187    美國Immunoway    50ug, 100ug    WB IHC ELISA    Human
Cytokeratin 5 Monoclonal Antibody    YM0188    美國Immunoway    50ug, 100ug    WB ELISA    Human
Cytokeratin 7 Monoclonal Antibody    YM0189    美國Immunoway    50ug, 100ug    WB FCM ELISA    Human
Cytokeratin 8 Monoclonal Antibody    YM0190    美國Immunoway    50ug, 100ug    WB IHC IF ELISA    Human
Cytokeratin (Pan) Monoclonal Antibody    YM0191    美國Immunoway    50ug, 100ug    WB IHC IF ELISA    Human
Cytokeratin (Pan) Monoclonal Antibody    YM0192    美國Immunoway    50ug, 100ug    IHC IF ELISA    Human
Daxx Monoclonal Antibody    YM0193    美國Immunoway    50ug, 100ug    WB IF FCM ELISA    Human
DDR1 Monoclonal Antibody    YM0194    美國Immunoway    50ug, 100ug    WB ELISA    Human
DDR2 Monoclonal Antibody    YM0195    美國Immunoway    50ug, 100ug    WB IHC IF ELISA    Human
DDX4 Monoclonal Antibody    YM0196    美國Immunoway    50ug, 100ug    WB IHC IF FCM ELISA    Human
Desmin Monoclonal Antibody    YM0197    美國Immunoway    50ug, 100ug    WB IHC ELISA    Human
Dkk-1 Monoclonal Antibody    YM0198    美國Immunoway    50ug, 100ug    WB ELISA    Human
Dkk-3 Monoclonal Antibody    YM0199    美國Immunoway    50ug, 100ug    WB FCM ELISA    Human, Monkey
DLK1 Monoclonal Antibody    YM0200    美國Immunoway    50ug, 100ug    WB IF FCM ELISA    Human
DNA-PKCS Monoclonal Antibody    YM0201    美國Immunoway    50ug, 100ug    WB IHC IF ELISA    Human
Dynamin I Monoclonal Antibody    YM0202    美國Immunoway    50ug, 100ug    WB IHC ELISA    Human
Dynamin II Monoclonal Antibody    YM0203    美國Immunoway    50ug, 100ug    WB IHC ELISA    Human
E2A Monoclonal Antibody    YM0204    美國Immunoway    50ug, 100ug    WB FCM ELISA    Human
E7 Monoclonal Antibody    YM0205    美國Immunoway    50ug, 100ug    WB ELISA    Human
E7 Monoclonal Antibody    YM0206    美國Immunoway    50ug, 100ug    WB IHC ELISA    Human
E-cadherin Monoclonal Antibody    YM0207    美國Immunoway    50ug, 100ug    WB IHC FCM ELISA    Human, Mouse, Monkey
E-cadherin Monoclonal Antibody    YM0208    美國Immunoway    50ug, 100ug    WB ELISA    Human, Mouse, Monkey
EF-2 Monoclonal Antibody    YM0209    美國Immunoway    50ug, 100ug    WB IHC IF ELISA    Human
EGF Monoclonal Antibody    YM0210    美國Immunoway    50ug, 100ug    WB IHC ELISA    Human
EGF Monoclonal Antibody    YM0211    美國Immunoway    50ug, 100ug    WB IF ELISA    Human
Egr-1 Monoclonal Antibody    YM0212    美國Immunoway    50ug, 100ug    WB ELISA    Human
EHMT1 Monoclonal Antibody    YM0213    美國Immunoway    50ug, 100ug    WB ELISA    Human
eIF4E Monoclonal Antibody    YM0214    美國Immunoway    50ug, 100ug    WB IHC IF FCM ELISA    Human
Elk1 Monoclonal Antibody    YM0215    美國Immunoway    50ug, 100ug    WB IHC ELISA    Human
Elk1 Monoclonal Antibody    YM0216    美國Immunoway    50ug, 100ug    WB ELISA    Human
Emt Monoclonal Antibody    YM0217    美國Immunoway    50ug, 100ug    WB ELISA    Human
Emt Monoclonal Antibody    YM0218    美國Immunoway    50ug, 100ug    WB IF FCM ELISA    Human
EP-CAM Monoclonal Antibody    YM0219    美國Immunoway    50ug, 100ug    WB IHC ELISA    Human
EphA1 Monoclonal Antibody    YM0220    美國Immunoway    50ug, 100ug    WB IHC ELISA    Human
EphA10 Monoclonal Antibody    YM0221    美國Immunoway    50ug, 100ug    WB ELISA    Human
EphA2 Monoclonal Antibody    YM0222    美國Immunoway    50ug, 100ug    WB IHC ELISA    Human