1,5-Anhydroglucitol (1,5-AG) ELISA Kit

Antigen: 1,5-Anhydroglucitol (1,5-AG)

Reaktivität: Kaninchen

Applikation: ELISA

Produktnummer: ABIN774194

Produktbeschreibung

Produktmerkmale: For the quantitative determination of rabbit 1,5-AG concentrations in serum, plasma,cell culture supernates and tissue homogenate.

Weitere Bezeichnung: 1,5-anhydroglucitol

Proben: Serum, Plasma, Cell Culture Supernatant, Tissue Homogenate

Kommentare:

Special Features of this Kit:

Stable enzyme
The enzyme in our kits is highly stable due to a unique protective agent.

Prediluted standards
The concentration of the standards is critical for an accurate standard curve. Therefore this kit contains serial dilutions of the standards rather than a concentrated standard solution, thereby avoiding pipetting or calculation errors. Quality tests assure that the concentration of the standards is accurate, thereby ensuring a reliable standard curve.

Storage
All kit contents can be stored at 2-8 degree. From a user perspective this is more convenient when it comes to transport and storage than a requirement for freezing some reagents, as common for many other ELISA kits.

Reliability
Thanks to just one incubation- and washing step the Elisa experiment can be performed within two hours. Compared to the traditional Elisa method, fewer handling steps reduce errors and deliver more consistent results. Thorough and regular tests of the system guarantee a great intra and inter assay reliability and ensure a low coefficient of variation.

Anwendungen

Aufbereitung der Reagenzien: 1. Bring all kit components and samples to room temperature (18-25 ℃) before use.
2. Dispense 10 uL of LYSIS BUFFER SOLUTION into 100 uL specimens, mix and stand for one hour (The proportion of LYSIS BUFFER and Specimens should be no less than 1:10). (NOTE: This step is required when the sample is cell culture fluid & body fluid & tissue homogenate; if the sample is serum or blood plasma, then this step should be skipped.)
3. Wash Solution - Dilute 10 mL of Wash Solution concentrate (100×) with 990 mL of deionized or distilled water to prepare 1000 mL of Wash Solution (1×).

Probennahme: Serum: Use a serum separator tube (SST) and allow samples to clot for 30 minutes before a centrifugation for 15 minutes at approximately 1000 x g. Remove serum and perform the assay immediately or aliquot and store samples at -20 °C or -80°C.
Plasma: Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 1000 x g at 2-8°C within 30 minutes of collection. Store samples at -20°C or -80°C.Avoid repeated freeze-thaw cycles.
Cell culture fluid and other biological fluids: Remove particulates by centrifugation and assay immediately or aliquot and store samples at -20°C or -80°C. Avoid repeated freeze-thaw cycles.
NOTE: Serum, plasma, and cell culture fluid samples to be used within 7 days may be stored at 2-8°C, otherwise samples must be stored at -20°C(≤2months) or -80°C(≤6months) to avoid loss of bioactivity and contamination. Avoid freeze-thaw cycles. When performing the assay, warm up samples to room temperature slowly. DO NOT USE HEAT-TREATED SAMPLES.

Aufbereitung der Proben: 1. The manufacturer is only responsible for the kit itself, but not for the samples consumed during the assay. The user should calculate the possible amount of the samples used in the whole test. Please reserve sufficient amount of samples in advance.
2. 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.
3. If the samples are not indicated in the manual, a preliminary experiment to determine the validity of the kit is necessary.
4. Owing to the possibility of mismatching between antigen from other resource and antibody used in our kits (e.g., antibody targets conformational epitope rather than linear epitope), some native or recombinant proteins from other manufacturers may not be recognized by our products.
5. Influenced by the factors including cell viability, cell number and also sampling time, samples from cell culture supernatant may not be detected by the kit.
6. Fresh samples without long time storage is recommended for the test. Otherwise, protein degradation and denaturalization may occur in those samples and finally lead to wrong results.

Ergebnisberechnung: 1. This standard curve is used to determine the amount of an unknown sample. Construct a standard curve by plotting the average O.D. (450 nm) for each standard on the vertical (Y) axis against the concentration on the horizontal (X) axis, and draw a best fit curve through the points on the graph.
2. First, calculate the mean O.D. value for each standard and sample. All O.D. values, are subtracted by the mean value of the blank control before result interpretation. Construct the standard curve using graph paper or statistical software.
3. To determine the amount in each sample, first locate the O.D. value on the Y-axis and extend a horizontal line to the standard curve. At the point of intersection, draw a vertical line to the X-axis and read the corresponding concentration.
4. Any variation in operator, pipetting and washing technique, incubation time or temperature, and kit age can cause variation in result. Each user should obtain their own standard curve.

Applikationshinweise: 1. When not in use, kit components should be refrigerated. All reagents should be warmed to room temperature before use.
2. Microtiter plates should be allowed to come to room temperature before opening the foil bags. Once the desired number of strips has been removed, immediately reseal the bag and store at 2-8°C to maintain plate integrity.
3. Samples should be collected in pyrogen/endotoxin-free tubes.
4. Samples should be frozen if not analyzed shortly after collection. Avoid multiple freeze-thaw cycles of frozen samples. Thaw completely and mix well prior to analysis.
5. When possible, avoid use of badly hemolyzed or lipemic sera. If large amounts of particulate matter are present, centrifuge or filter prior to analysis.
6. It is recommended that all standards, controls and samples be run in duplicate.
7. When pipetting reagents, maintain a consistent order of addition from well-to-well. This ensures equal incubation times for all wells.
8. Cover or cap all reagents when not in use.
9. Do not mix or interchange different reagent lots from various kit lots.
10. Do not use reagents after the kit expiration date.
11. Read absorbances within 2 hours of assay completion.
12. The provided controls should be run with every assay. If control values fall outside pre-established ranges, the accuracy of the assay is suspect.
13. All residual wash liquid must be drained from the wells by efficient aspiration or by decantation followed by tapping the plate forcefully on absorbent paper. Never insert absorbent paper directly into the wells.
14. Because Stabilized Chromogen is light sensitive, avoid prolonged exposure to light. Also avoid contact between Stabilized Chromogen and metal, otherwise color may develop.
15. Incomplete washing will adversely affect the test outcome. All washing must be performed with Wash Solution provided.
16. Washing can be performed manually as follows: completely aspirate the liquid from all wells by gently lowering an aspiration tip into the bottom of each well. Take care not to scratch the inside of the well.
17. After aspiration, fill the wells with at least 0.4 mL of diluted wash solution. Let soak for 15 to 30 seconds, and then aspirate the liquid. Repeat as directed under ASSAY PROCEDURE. After the washing procedure, the plate is inverted and tapped dry on absorbent tissue.
18. Alternatively, the wash solution may be put into a squirt bottle. If a squirt bottle is used, flood the plate with wash buffer, completely filling all wells.
19. If using an automated washer, the operating instructions for washing equipment should be carefully followed.
20. Assay Procedure Preliminary notes: Do not mix reagents from different lots. It is recommended that assays be performed in duplicate. Standards and samples must be assayed at the same time. Avoid exposing the substrate to direct sunlight.

Handhabung: 1. This kit contains materials with small quantities of sodium azide. Sodium azide reacts with lead and copper plumbing to form explosive metal azides. Upon disposal, flush drains with a large volume of water to prevent azide accumulation. Avoid ingestion and contact with eyes, skin and mucous membranes. In case of contact, rinse affected area with plenty of water. Observe all federal, state and local regulations for disposal.
2. All blood components and biological materials should be handled as potentially hazardous. Follow universal precautions as established by the Centers for Disease Control and Prevention and by the Occupational Safety and Health Administration when handling and disposing of infectious agents.

Benötigtes Material: 1. Microplate reader capable of measuring absorbance at 450 nm.
2. Pipettes and pipette tips.
3. 100 mL and 1 liter graduated cylinders.
4. Calibrated adjustable precision pipettes, preferably with disposable plastic tips. (A manifold multi-channel pipette is desirable for large assays.)
5. Absorbent paper.
6. 37°C incubator.
7. Distilled or deionized water.
8. Data analysis and graphing software. Graph paper: linear (Cartesian), log-log or semi-log, or log-logit as desired.
9. Tubes to prepare standard or sample dilutions.

Beschränkungen: Nur für Forschungszwecke einsetzbar