Chicken Salmonella Enteritidis IgY ELISA Kit | Creative Diagnostics DEIABL37 product information

product summary

company name :

Creative Diagnostics

product type :

ELISA/assay

product name :

Chicken Salmonella Enteritidis IgY ELISA Kit

catalog :

DEIABL37

quantity :

96T

more info or order :

Creative Diagnostics product webpage

product information

Catalog :

DEIABL37

proudctName :

Chicken Salmonella Enteritidis IgY ELISA Kit

common :

Salmonella

Short Name :

Chicken Salmonella Enteritidis IgY ELISA Kit

Detection Method :

sELISA

Common name :

Salmonella

Species :

Chicken

Application :

Quantitative

Sample :

Serum, Plasma and other biological fluids

Size :

96T

Storage :

All unopened components of this kit are stable at 2-8°C until the kit's expiration date.

Species Reactivity :

Chicken

Intended Use :

The Chicken Salmonella Enteritidis IgY (CSE-IgY) ELISA Kit is to be used for the vitroquantitative determination of Chicken CSE-IgY in Serum, Plasma and other biological fluids. The Kit is intended for research use only, not for diagnostic or therapeuticprocedure

Principle Of The Test :

The kit assay Chicken CSE-IgY level in the Samples, use Purified Chicken CSE-IgY antigen to coat microtiter plate wells, make solid-phase antigen, then add Samples (Containing Chicken CSE-IgY) to wells, combined Chicken CSE-IgY antigen which with HRP labeled, become antigen - antibody - enzyme-antigen complex, after washing completely, add TMB substrate solution, TMB substrate becomes blue color at HRP enzyme-catalyzed, reaction is terminated by the addition of a sulphuric acid solutionand the color change (yellow) is measured spectrophotometrically at a wavelength of 450 nm. The concentration of Chicken CSE-IgY in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Reagents And Materials Provided :

Hints: 1. All the components in the kit should be stored up to 6 months at 2-8°C,andshould be kept according to the labels on vials. (Storage:2-8°C Validity: 6 months) 2. The kit takes out from the refrigeration environment should be balanced 15-30 minutes in the room temperature then use. 3. The kit contain sufficient materials to run ELISAs on 96 or 48 microplates. Specific vial volume of each component may vary. Please check carefully if all 4. Washing Buffer will present Crystallization separation, and it can be heated inthe water helping to dissolve,which does not affect the result. 5. Closure plate membrane only limits the disposable use, in order to avoid the overlapping pollution 6. Add sample within 5 min. Please keeping the TMB Substrate evade the light preservation. 7. Upon receipt, foil pouch around the plate should be vacuum-sealed. 8. Please avoid repeated freeze-thaw cycles and do not mix reagents from different kits unless they have the same lot numbers. 9. After use remaining reagents should be returned to cold storage (2°to 8°C) immediately. Besides, please return the unused wells to the foil pouch containing the desiccant pack, and reseal along entire edge of zip-seal. 10. Do not use components beyond the expiration date.Expiry of the kit components can only be guaranteed if the components are stored properly, and if, in case of repeated use of one component, this reagent is not contaminated by the first handling. Expiry of the kit and reagents is stated on kit labels. 11. It is highly recommended to use the remaining reagents within 1 month provided, this is within the expiration date of the kit. 12. Any irregularities to aforementioned conditions may influence plate performance in the assay.

Materials Required But Not Supplied :

1. Microplate reader (450 nm detection wavelength filter, 570 nm or 630 nm correction wavelength filters) 2. Beakers, flasks, cylinders necessary for preparation of reagents 3. Clean benches, Incubator (37°C), Refrigerators (4°C, -20°C), Low Temperature Centrifuge 4. High-precision single-channel and multi-channel Pipette and disposable Tips. 5. Polypropylene tubes for diluting and aliquoting Standards if needed 6. Distilled water or deionized water 7. Absorbent paper for blotting the microtiter plate 8. Automated or manual microplate washer

Specimen Collection And Preparation :

Serum: Allow samples to clot in a serum separator tube for two hours at roomtemperature or overnight at 4°C. Centrifuge at approximately 2000 -3000 rpm for 15-20 min. Analyze the serum immediately or aliquot and store frozen at -20°C or -80°C.Avoid repeated freeze/thaw cycles. Plasma: Collect plasma using heparin, EDTA, citrate as an anticoagulant. Centrifugeat approximately 2000 -3000rpm for 15-20 min. Analyze immediately or aliquot andstore frozen at -20°C or -80°C. Avoid repeated freeze/thaw cycles. Tissue homogenates: After cutting samples, check the weight, and add PBS (PH7.2-7.4), rapidly freeze with Liquid Nitrogen; maintain samples at 2-8°C after melting,add PBS (PH7.4), homogenized in ice water. Centrifuge at approximately 2000-5000 rpm for 20 min. Having the Supernatant and discarded following precipitation.Analyze immediately or aliquot and store frozen at -20°C or -80°C. Avoid repeated freeze/thaw cycles. Cell culture supernatant: If detec the secretory components, please collect withasterile container, then centrifuge at approximately 2000-3000rpm for 20 min ,havingthe Supernatant; if detec the Intracellular components, please dilute the Cell suspension with PBS (PH7.2-7.4),and the concentration reached 1000000 / ml, repeated freeze-thaw cycles or other method, let the cells release the Intracellular components, centrifuge at approximately 2000-3000 rpm for 20, having the Supernatant. If precipitation appeared, centrifugal again. Analyze immediately or aliquot and store frozen at -20°C or -80°C. Avoid repeated freeze/thaw cycles. Urine ,Ascites, Cerebrospinal fluid and other body fluids: Collect with a sterile container,remove precipitation by centrifugation at approximately 2000-3000 rpm for 20min. Analyze immediately or aliquot and store frozen at -20°C or -80°C. Avoidrepeated freeze/thaw cycles. Hints: 1. Other biological samples might be suitable for use in the assay,please inquire our Tech Support. 2. Avoid repeated freeze-thaw cycles. Prior to assay, the frozen sample should be brought to room temperature slowly and mixed gently. 3. Samples to be used within 24 hours may be stored at 4°C, otherwise samples must be stored at -20°C (≤3 month) or -80°C (≤6 months) to avoid loss of bioactivity and contamination. 4. Samples containing a visible precipitate must be clarified prior to use in the assay. Do not use grossly hemolyzed or lipemic specimens. 5. The collection and extraction of Sample, recommend the customer to read therelevant literiture. 6. Can not detect the sample containing NaN 3 , because NaN 3 inhibits the activity of HRP. Sample Dilution Guideline User needs to estimate the concentration of the target protein in the samples andselect proper dilution factor so that the diluted target protein concentration fallsnear the middle of the linear regime in the standard curve.

Assay Procedure :

1. Dilution and addition of standard: Ten wells are designated as standard sample wells on the enzyme-labeled plate. 100μL of standard sample is added to the first and second wells, followed by the addition of 50μL of standard sample diluent to the first and second wells, and mixing. Subsequently, 100μL is taken from each of the first and second wells and added to the third and fourth wells, followed by the addition of 50μL of standard sample diluent to the third and fourth wells, and mixing. Then, 50μL is removed from each of the third and fourth wells, and 50μL is added to the fifth and sixth wells respectively, followed by the addition of 50 μL of standard sample diluent to the fifth and sixth wells, and mixing. After mixing, 50μL is taken from each of the fifth and sixth wells and added to the seventh and eighth wells, followed by the addition of 50 μL of standard sample diluent to the seventh and eighth wells, and after mixing, 50μL is taken from the seventh and eighth wells and added to the ninth and tenth wells, followed by the addition of 50μL of standard sample diluent. After mixing, 50μL is taken from the ninth and tenth wells and discarded. (After dilution, the amount of sample added to each well is 50 μL, with concentrations of 7.2 ng/mL, 4.8 ng/mL, 2.4 ng/mL, 1.2 ng/mL, 0.6 ng/mL, respectively.) 2. Set Standard well, Blank well (Please do not add Sample and HRP-Conjugate reagent to the Blank comparison wells,other each step operationis same), and Testing sample well. 3. Add Standards and Samples: Aliquot the prepared standard solutions 50 μl into each Standard well; add Sample Diluent 40 μL to Testing sample well, then add testing sample 10 μL (Sample final dilution is 5-fold; Please select proper dilution factor, or no dilution test directly). Please add Sample to the bottom of pre-coated well , do not touch the well wall as far as possible, and mix gently. 4. Incubate: After closing the plate with Closure plate membrane ,incubate for 30 min at 37°C. 5. Prepare the Washing Buffer: 30-fold Wash Solution, diluted 30-fold with Distilled water. 6. Washing: Uncover Closure plate membrane, discard Liquid, dry by swing, addWashing Buffer fill to each well, still for 30s then drain, repeat 5 times, dry by pat. (Avaiblable for Automated or manual microplate washer) 7. Add enzyme: Add HRP-Conjugate Reagent 50 μL to each well, except the Blank well. 8. Incubate: After closing the plate with Closure plate membrane ,incubate for 30 min at 37°C. 9. Washing: Uncover Closure plate membrane, discard Liquid, dry by swing, add Washing Buffer fill to each well, still for 30s then drain, repeat 5 times, dry by pat. (Avaiblable for Automated or manual microplate washer) 10. Color: Add TMB Chromogen Solution A 50 μL and then add TMB Chromogen Solution B 50 μL to each well, mix gently, evade the light preservation for 15 min at 37°C. 11. Stop the Reaction: Add Stop Solution 50 μL to each well, to stop the reaction (the blue color change to yellow color immediately). 12. Assay: Set the OD of Blank well as zero, read absorbance at 450 nm after adding Stop Solution within 15 min. The judgment of result must take the OD of Microplate reader as a standard, when use the dual-wavelength to assay, reference wavelength is 630 nm.

Calculation :

Take the Standard density as the horizontal, the OD value for the vertical, obtaintheStandard Curve, then find out the corresponding density according to the sample OD value, and 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 sampleactual density.

Performance Characteristics :

Intra-Assay: CV ≤ 15% Inter-Assay: CV ≤ 15% Correlation coefficient (R) of linear regression of the samples is more than 0.92.

Detection Range :

0.15 - 10 ng/mL

Detection Limit :

< 1.2 ng/mL

applicatons :

Contents of Kit :

Tetramethylbenzidine :